Chikungunya virus rna vaccines

ABSTRACT

The disclosure relates to tropical diseases such as viral mosquito borne illnesses and the treatment thereof. The invention includes ribonucleic acid vaccines and combination vaccines, as well as methods of using the vaccines and compositions comprising the vaccines for treating and preventing tropical disease.

RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No. 15/674,591, filed Aug. 11, 2017, which is a continuation of international application number PCT/US2016/058324, filed Oct. 21, 2016, which claims the benefit under 35 U.S.C. § 119(e) of U.S. provisional application No. 62/244,937, filed Oct. 22, 2015, U.S. provisional application No. 62/247,347, filed Oct. 28, 2015, U.S. provisional application No. 62/244,814, filed Oct. 22, 2015, U.S. provisional application No. 62/247,390, filed Oct. 28, 2015, U.S. provisional application No. 62/245,207, filed Oct. 22, 2015, U.S. provisional application No. 62/247,445, filed Oct. 28, 2015, U.S. provisional application No. 62/244,950, filed Oct. 22, 2015, U.S. provisional application No. 62/247,595, filed Oct. 28, 2015, U.S. provisional application No. 62/351,255, filed Jun. 16, 2016 and U.S. provisional application No. 62/245,031, filed Oct. 22, 2015, each of which is incorporated by reference herein in its entirety.

BACKGROUND

Insects such as mosquitoes cause significant human suffering by transmission of infectious disease to humans. The infections carried by mosquitoes afflict humans, as well as companion animals such as dogs and horses. Infectious agents transmitted by mosquitos cause illnesses such as encephalitis, Chikungunya, yellow fever, West Nile fever, malaria, and Dengue. The transmission of diseases associated with mosquito bites can be interrupted by killing the mosquitoes, isolating infected people from all mosquitoes while they are infectious or vaccinating the exposed population.

Deoxyribonucleic acid (DNA) vaccination is one technique used to stimulate humoral and cellular immune responses to foreign antigens, such as Malaria, JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and YFV antigens. The direct injection of genetically engineered DNA (e.g., naked plasmid DNA) into a living host results in a small number of its cells directly producing an antigen, resulting in a protective immunological response. With this technique, however, comes potential problems, including the possibility of insertional mutagenesis, which could lead to the activation of oncogenes or the inhibition of tumor suppressor genes.

SUMMARY

Provided herein are ribonucleic acid (RNA) vaccines that build on the knowledge that RNA (e.g., messenger RNA (mRNA)) can safely direct the body's cellular machinery to produce nearly any protein of interest, from native proteins to antibodies and other entirely novel protein constructs that can have therapeutic activity inside and outside of cells. The RNA (e.g., mRNA) vaccines of the present disclosure may be used to induce a balanced immune response against Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), Japanese Encephalitis Virus (JEV), West Nile Virus (WNV), Eastern Equine Encephalitis Virus (EEEV), Venezuelan Equine Encephalitis Virus (VEEV), Sindbis Virus (SINV), Chikungunya Virus (CHIKV), Dengue Virus (DENV), Zika Virus (ZIKV) and/or Yellow Fever Virus (YFV), comprising both cellular and humoral immunity, without risking the possibility of insertional mutagenesis, for example. Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV are referred to herein as “tropical diseases.” Thus, the terms “tropical disease vaccines” and “Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV” encompass Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale) RNA vaccines, JEV RNA vaccines, WNV RNA vaccines, EEEV RNA vaccines, SINV RNA vaccines, CHIKV RNA vaccines, DENV RNA vaccines, ZIKV RNA vaccines, YFV RNA vaccines, and combination vaccines comprising at least two (e.g., at least 3, 4, 5, 6, 7, 8 or 9) of any of the Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale) RNA vaccines, JEV RNA vaccines, WNV RNA vaccines, EEEV RNA vaccines, SINV RNA vaccines, CHIKV RNA vaccines, DENV RNA vaccines, ZIKV RNA vaccines, and YFV RNA vaccines.

The RNA (e.g., mRNA) vaccines may be utilized in various settings depending on the prevalence of the infection or the degree or level of unmet medical need. The RNA (e.g. mRNA) vaccines may be utilized to treat and/or prevent Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV of various genotypes, strains, and isolates. The RNA (e.g., mRNA) vaccines have superior properties in that they produce much larger antibody titers and responses earlier than commercially available anti-viral therapeutic treatments. While not wishing to be bound by theory, it is believed that the RNA (e.g., mRNA) vaccines, as mRNA polynucleotides, are better designed to produce the appropriate protein conformation upon translation, as the RNA (e.g., mRNA) vaccines co-opt natural cellular machinery. Unlike traditional vaccines, which are manufactured ex vivo and may trigger unwanted cellular responses, RNA (e.g., mRNA) vaccines are presented to the cellular system in a more native fashion.

Surprisingly, it has been shown that efficacy of mRNA vaccines can be significantly enhanced when combined with a flagellin adjuvant, in particular, when one or more antigen-encoding mRNA is combined with an mRNA encoding flagellin. RNA (e.g., mRNA) vaccines combined with the flagellin adjuvant (e.g., mRNA-encoded flagellin adjuvant) have superior properties in that they may produce much larger antibody titers and produce responses earlier than commercially available vaccine formulations.

Some embodiments of the present disclosure provide RNA (e.g., mRNA) vaccines that include at least one RNA (e.g., mRNA) polynucleotide having an open reading frame encoding at least one antigenic polypeptide or an immunogenic fragment thereof (e.g., an immunogenic fragment capable of inducing an immune response to the antigenic polypeptide) and at least one RNA (e.g., mRNA polynucleotide) having an open reading frame encoding a flagellin adjuvant.

In some embodiments, at least one flagellin polypeptide (e.g., encoded flagellin polypeptide) is a flagellin protein. In some embodiments, at least one flagellin polypeptide (e.g., encoded flagellin polypeptide) is an immunogenic flagellin fragment. In some embodiments, at least one flagellin polypeptide and at least one antigenic polypeptide are encoded by a single RNA (e.g., mRNA) polynucleotide. In other embodiments, at least one flagellin polypeptide and at least one antigenic polypeptide are each encoded by a different RNA polynucleotide.

In some embodiments at least one flagellin polypeptide has at least 80%, at least 85%, at least 90%, or at least 95% identity to a flagellin polypeptide having a sequence of SEQ ID NO: 420-422.

Provided herein, in some embodiments, is a ribonucleic acid (RNA) (e.g., mRNA) vaccine, comprising at least one (e.g., at least 2, 3, 4 or 5) RNA (e.g., mRNA) polynucleotide having an open reading frame encoding at least one (e.g., at least 2, 3, 4 or 5) Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide, or any combination of two or more of the foregoing antigenic polypeptides. Herein, use of the term “antigenic polypeptide” encompasses immunogenic fragments of the antigenic polypeptide (an immunogenic fragment that induces (or is capable of inducing) an immune response to Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV) unless otherwise stated.

Also provided herein, in some embodiments, is a RNA (e.g., mRNA) vaccine comprising at least one (e.g., at least 2, 3, 4 or 5) RNA polynucleotide having an open reading frame encoding at least one (e.g., at least 2, 3, 4 or 5) Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide or an immunogenic fragment thereof, linked to a signal peptide.

Further provided herein, in some embodiments, is a nucleic acid (e.g., DNA) encoding at least one (e.g., at least 2, 3, 4 or 5) Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV RNA (e.g., mRNA) polynucleotide.

Further still, provided herein, in some embodiments, is a method of inducing an immune response in a subject, the method comprising administering to the subject a vaccine comprising at least one (e.g., at least 2, 3, 4 or 5) RNA (e.g., mRNA) polynucleotide having an open reading frame encoding at least one (e.g., at least 2, 3, 4 or 5) Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide, or any combination of two or more of the foregoing antigenic polypeptides.

Malaria

Some embodiments of the present disclosure provide Malaria vaccines that include at least one ribonucleic acid (RNA) polynucleotide having an open reading frame encoding at least one Plasmodium (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale) antigenic polypeptide or an immunogenic fragment thereof (e.g., an immunogenic fragment capable of raising an immune response to Plasmodium).

In some embodiments, the antigenic polypeptide is a circumsporozoite (CS) protein or an immunogenic fragment thereof (e.g., capable of raising an immune response against Plasmodium).

In some embodiments, the CS protein or fragment is fused to the surface antigen from hepatitis B (HBsAg). In some embodiments, the CS protein or fragment is in the form of a hybrid protein comprising substantially all the C-terminal portion of the CS protein of Plasmodium, four or more tandem repeats of the CS protein immunodominant region, and the surface antigen from hepatitis B (HBsAg). In some embodiments, the hybrid protein comprises a sequence of CS protein of Plasmodium falciparum substantially as corresponding to amino acids 207-395 of P. falciparum NF54 strain 3D7 clone CS protein fused in frame via a linear linker to the N-terminal of HBsAg (Ballou W R et al. Am J Trop Med Hyg 2004; 71(2_suppl):239-247, incorporated herein by reference).

In some embodiments, the hybrid protein is RTS. In some embodiments, the RTS is in the form of mixed particles RTS,S. In some embodiments, the amount of RTS,S is 25 μg or 50 μg per dose.

In some embodiments, the antigenic polypeptide is liver stage antigen 1 (LSA1) or an immunogenic fragment thereof. In some embodiments, the antigenic polypeptide is LSA-NRC.

In some embodiments, the antigenic polypeptide is merozoite surface protein-1 (MSP1) or an immunogenic fragment thereof.

In some embodiments, the antigenic polypeptide is apical membrane antigen 1 (AMA1) or an immunogenic fragment thereof.

In some embodiments, the antigenic polypeptide is thrombospondin related adhesive protein (TRAP) or an immunogenic fragment thereof.

In some embodiments, at least one RNA polynucleotide is encoded by at least one nucleic acid sequence identified by any one of SEQ ID NO: 1-6 (Table 1) and homologs having at least 80% identity with a nucleic acid sequence identified by any one of SEQ ID NO: 1-6 (Table 1). In some embodiments, at least one RNA polynucleotide is encoded by at least one nucleic acid sequence identified by any one of SEQ ID NO: 1-6 (Table 1) and homologs having at least 90% (90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.8% or 99.9%) identity with a nucleic acid sequence identified by any one of SEQ ID NO: 1-6 (Table 1). In some embodiments, at least one RNA polynucleotide is encoded by at least one fragment of a nucleic acid sequence identified by any one of SEQ ID NO: 1-6 (Table 1).

In some embodiments, at least one RNA polynucleotide comprises at least one nucleic acid sequence identified by any one of SEQ ID NO: 7-12 (Table 1) and homologs having at least 80% identity with a nucleic acid sequence identified by any one of SEQ ID NO: 7-12 (Table 1). In some embodiments, at least one RNA polynucleotide comprises at least one nucleic acid sequence identified by any one of SEQ ID NO: 7-12 (Table 1) and homologs having at least 90% (90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.8% or 99.9%) identity with a nucleic acid sequence identified by any one of SEQ ID NO: 7-12 (Table 1). In some embodiments, at least one RNA polynucleotide comprises at least one fragment of a nucleic acid sequence identified by any one of SEQ ID NO: 7-12 (Table 1).

In some embodiments, the at least one RNA polynucleotide encodes at least one antigenic polypeptide having a sequence identified by any one of SEQ ID NO: 13-17 (Table 2 and 3). In some embodiments, the at least one RNA polynucleotide encodes at least one protein variant having at least 95% identity to an antigenic polypeptide having a sequence identified by any one of SEQ ID NO: 13-17 (Table 2 and 3). In some embodiments, at least one antigenic polypeptide has an amino acid sequence identified by any one of SEQ ID NO: 13-17 (Table 2 and 3). In some embodiments, at least one antigenic polypeptide has at least 95% identity to an antigenic polypeptide having a sequence identified by any one of SEQ ID NO: 13-17 (Table 2 and 3).

Japanese Encephalitis Virus (JEV)

Some embodiments of the present disclosure provide JEV vaccines that include at least one ribonucleic acid (RNA) polynucleotide having an open reading frame encoding at least one JEV antigenic polypeptide or an immunogenic fragment thereof (e.g., an immunogenic fragment capable of inducing an immune response to JEV).

In some embodiments, at least one antigenic polypeptide is JEV E protein, JEV Es, JEV prM, JEV capsid, JEV NS1, JEV prM and E polyprotein (prME) or an immunogenic fragment thereof. In some embodiments, at least one antigenic polypeptide has at least 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98% or 99% sequence identity to JEV E protein, JEV Es, JEV prM, JEV capsid, prME or JEV NS1.

In some embodiments, at least one RNA polynucleotide is encoded by at least one nucleic acid sequence identified by any one of SEQ ID NO: 18-19 (Table 4) and homologs having at least 80% identity with a nucleic acid sequence identified by any one of SEQ ID NO: 18-19 (Table 4). In some embodiments, at least one RNA polynucleotide is encoded by at least one nucleic acid sequence identified by any one of SEQ ID NO: 18-19 (Table 4) and homologs having at least 90% (90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.8% or 99.9%) identity with a nucleic acid sequence identified by any one of SEQ ID NO: 18-19 (Table 4). In some embodiments, at least one RNA polynucleotide is encoded by at least one fragment of a nucleic acid sequence identified by any one of SEQ ID NO: 18-19 (Table 4).

In some embodiments, at least one RNA polynucleotide comprises at least one nucleic acid sequence identified by any one of SEQ ID NO: 20-21 (Table 4) and homologs having at least 80% identity with a nucleic acid sequence identified by any one of SEQ ID NO: 20-21 (Table 4). In some embodiments, at least one RNA polynucleotide comprises at least one nucleic acid sequence identified by any one of SEQ ID NO: 20-21 (Table 4) and homologs having at least 90% (90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.8% or 99.9%) identity with a nucleic acid sequence identified by any one of SEQ ID NO: 20-21 (Table 4). In some embodiments, at least one RNA polynucleotide comprises at least one fragment of a nucleic acid sequence identified by any one of SEQ ID NO: 20-21 (Table 4).

In some embodiments, the at least one RNA polynucleotide encodes at least one antigenic polypeptide having a sequence identified by any one of SEQ ID NO: 22-29 (Table 5 and 6). In some embodiments, the at least one RNA polynucleotide encodes at least one protein variant having at least 95% identity to an antigenic polypeptide having a sequence identified by any one of SEQ ID NO: 22-29 (Table 5 and 6). In some embodiments, at least one antigenic polypeptide has an amino acid sequence identified by any one of SEQ ID NO: 22-29 (Table 5 and 6). In some embodiments, at least one antigenic polypeptide has at least 95% identity to an antigenic polypeptide having a sequence identified by any one of SEQ ID NO: 22-29 (Table 5 and 6).

West Nile Virus (WNV), Eastern Equine Encephalitis (EEEV), Venezuelan Equine Encephalitis Virus (VEEV), and Sindbis Virus (SINV)

Some embodiments of the present disclosure provide combination vaccines comprising one or more RNA (e.g., mRNA) polynucleotides. The RNA polynucleotide(s) encode one or more Arbovirus antigens and/or one or more Alphavirus antigens, on either the same polynucleotide or different polynucleotides. RNA polynucleotides featured in the vaccines of the present invention can encode one antigen or can encode more than one antigen, e.g., several antigens (for example, polycistronic RNAs).

In some embodiments, at least one RNA polynucleotide is encoded by at least one nucleic acid sequence identified by any one of SEQ ID NO: 30-34, 48-49, 55-56 (Table 9, 12, 15) and homologs having at least 80% identity with a nucleic acid sequence identified by any one of SEQ ID NO: 30-34, 48-49, 55-56 (Table 9, 12, 15). In some embodiments, at least one RNA polynucleotide is encoded by at least one nucleic acid sequence identified by any one of SEQ ID NO: 30-34, 48-49, 55-56 (Table 9, 12, 15) and homologs having at least 90% (90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.8% or 99.9%) identity with a nucleic acid sequence identified by any one of SEQ ID NO: 30-34, 48-49, 55-56 (Table 9, 12, 15). In some embodiments, at least one RNA polynucleotide is encoded by at least one fragment of a nucleic acid sequence identified by any one of SEQ ID NO: 30-34, 48-49, 55-56 (Table 9, 12, 15).

In some embodiments, at least one RNA polynucleotide comprises at least one nucleic acid sequence identified by any one of SEQ ID NO: 35-39, 50-51, 57-58 (Table 9, 12, 15) and homologs having at least 80% identity with a nucleic acid sequence identified by any one of SEQ ID NO: 35-39, 50-51, 57-58 (Table 9, 12, 15). In some embodiments, at least one RNA polynucleotide comprises at least one nucleic acid sequence identified by any one of SEQ ID NO: 35-39, 50-51, 57-58 (Table 9, 12, 15) and homologs having at least 90% (90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.8% or 99.9%) identity with a nucleic acid sequence identified by any one of SEQ ID NO: 35-39, 50-51, 57-58 (Table 9, 12, 15). In some embodiments, at least one RNA polynucleotide comprises at least one fragment of a nucleic acid sequence identified by any one of SEQ ID NO: 35-39, 50-51, 57-58 (Table 9, 12, 15).

In some embodiments, the at least one RNA polynucleotide encodes at least one antigenic polypeptide having a sequence identified by any one of SEQ ID NO: 44-47, 52-54, 59-64 (Table 10, 11, 13, 14, 16, 17 and 18). In some embodiments, the at least one RNA polynucleotide encodes at least one protein variant having at least 95% identity to an antigenic polypeptide having a sequence identified by any one of SEQ ID NO: 44-47, 52-54, 59-64 (Table 10, 11, 13, 14, 16, 17 and 18). In some embodiments, at least one antigenic polypeptide has an amino acid sequence identified by any one of SEQ ID NO: 44-47, 52-54, 59-64 (Table 10, 11, 13, 14, 16, 17 and 18). In some embodiments, at least one antigenic polypeptide has at least 95% identity to an antigenic polypeptide having a sequence identified by any one of SEQ ID NO: 44-47, 52-54, 59-64 (Table 10, 11, 13, 14, 16, 17 and 18).

Yellow Fever Virus (YFV)

Yellow fever is an acute viral haemorrhagic disease transmitted by infected mosquitoes. The “yellow” in the name refers to the jaundice that affects some patients. Symptoms of yellow fever include fever, headache, jaundice, muscle pain, nausea, vomiting and fatigue. A small proportion of patients who contract the virus develop severe symptoms and approximately half of those die within 7 to 10 days. Yellow fever virus (YFV) is endemic in tropical areas of Africa and Central and South America. Large epidemics of yellow fever occur when infected people introduce the virus into heavily populated areas with high mosquito density and where most people have little or no immunity, due to lack of vaccination. In these conditions, infected mosquitoes transmit the virus from person to person. Since the launch of the Yellow Fever Initiative in 2006, significant progress in combating the disease has been made in West Africa and more than 105 million people have been vaccinated in mass campaigns using an attenuated live vaccine. Nonetheless, this vaccine can cause yellow fever vaccine-associated viscerotropic disease as well as yellow fever vaccine-associated neurotropic disease, each of which can be fatal.

Some embodiments of the present disclosure provide Yellow fever virus (YFV) vaccines that include at least one ribonucleic acid (RNA) polynucleotide (e.g., mRNA polynucleotide) having an open reading frame encoding at least one YFV antigenic polypeptide or an immunogenic fragment thereof (e.g., an immunogenic fragment capable of inducing an immune response to YFV).

In some embodiments, the at least one antigenic polypeptide is a YFV polyprotein.

In some embodiments, the at least one antigenic polypeptide is a YFV capsid protein, a YFV premembrane/membrane protein, a YFV envelope protein, a YFV non-structural protein 1, a YFV non-structural protein 2A, a YFV non-structural protein 2B, a YFV non-structural protein 3, a YFV non-structural protein 4A, a YFV non-structural protein 4B, or a YFV non-structural protein 5.

In some embodiments, the at least one antigenic polypeptide is a YFV capsid protein or an immunogenic fragment thereof, a YFV premembrane/membrane protein or an immunogenic fragment thereof, and a YFV envelope protein or an immunogenic fragment thereof.

In some embodiments, the at least one antigenic polypeptide is a YFV capsid protein or an immunogenic fragment thereof and a YFV premembrane/membrane protein or an immunogenic fragment thereof.

In some embodiments, the at least one antigenic polypeptide is a YFV capsid protein or an immunogenic fragment thereof and a YFV envelope protein or an immunogenic fragment thereof.

In some embodiments, at least one antigenic polypeptide is a YFV premembrane/membrane protein or an immunogenic fragment thereof and a YFV envelope protein or an immunogenic fragment thereof.

In some embodiments, the at least one antigenic polypeptide further comprises any one or more of a YFV non-structural protein 1, 2A, 2B, 3, 4A, 4B or 5.

In some embodiments, at least one RNA polynucleotide is encoded by at least one nucleic acid sequence identified by any one of SEQ ID NO: 65-80 (Table 21) and homologs having at least 80% identity with a nucleic acid sequence identified by any one of SEQ ID NO: 65-80 (Table 21). In some embodiments, at least one RNA polynucleotide is encoded by at least one nucleic acid sequence identified by any one of SEQ ID NO: 65-80 (Table 21) and homologs having at least 90% (90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.8% or 99.9%) identity with a nucleic acid sequence identified by any one of SEQ ID NO: 65-80 (Table 21). In some embodiments, at least one RNA polynucleotide is encoded by at least one fragment of a nucleic acid sequence identified by any one of SEQ ID NO: 65-80 (Table 21).

In some embodiments, at least one RNA polynucleotide comprises at least one nucleic acid sequence identified by any one of SEQ ID NO: 81-96 (Table 21) and homologs having at least 80% identity with a nucleic acid sequence identified by any one of SEQ ID NO: 81-96 (Table 21). In some embodiments, at least one RNA polynucleotide comprises at least one nucleic acid sequence identified by any one of SEQ ID NO: 81-96 (Table 21) and homologs having at least 90% (90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.8% or 99.9%) identity with a nucleic acid sequence identified by any one of SEQ ID NO: 81-96 (Table 21). In some embodiments, at least one RNA polynucleotide comprises at least one fragment of a nucleic acid sequence identified by any one of SEQ ID NO: 81-96 (Table 21).

In some embodiments, the at least one RNA polynucleotide encodes at least one antigenic polypeptide having a sequence identified by any one of SEQ ID NO: 97-117 (Table 22). In some embodiments, the at least one RNA polynucleotide encodes at least one protein variant having at least 95% identity to an antigenic polypeptide having a sequence identified by any one of SEQ ID NO: 97-117 (Table 22). In some embodiments, at least one antigenic polypeptide has an amino acid sequence identified by any one of SEQ ID NO: 97-117 (Table 22). In some embodiments, at least one antigenic polypeptide has at least 95% identity to an antigenic polypeptide having a sequence identified by any one of SEQ ID NO: 97-117 (Table 22).

Zika Virus (ZIKV)

Zika virus (ZIKV) is a member of the Flaviviridae virus family and the flavivirus genus. In humans, it causes a disease known as Zika fever. It is related to dengue, yellow fever, West Nile and Japanese encephalitis, viruses that are also members of the virus family Flaviviridae. ZIKV is spread to people through mosquito bites. The most common symptoms of ZIKV disease (Zika) are fever, rash, joint pain, and red eye. The illness is usually mild with symptoms lasting from several days to a week. There is no vaccine to prevent, or medicine to treat, Zika virus.

Some embodiments of the present disclosure provide Zika virus (ZIKV) vaccines that include at least one ribonucleic acid (RNA) polynucleotide (e.g., mRNA polynucleotide) having an open reading frame encoding at least one ZIKV antigenic polypeptide or an immunogenic fragment thereof (e.g., an immunogenic fragment capable of inducing an immune response to ZIKV).

In some embodiments, at least one antigenic polypeptide is a ZIKV polyprotein. In some embodiments, at least one antigenic polypeptide is a ZIKV structural polyprotein. In some embodiments, at least one antigenic polypeptide is a ZIKV nonstructural polyprotein.

In some embodiments, at least one antigenic polypeptide is a ZIKV capsid protein, a ZIKV premembrane/membrane protein, a ZIKV envelope protein, a ZIKV non-structural protein 1, a ZIKV non-structural protein 2A, a ZIKV non-structural protein 2B, a ZIKV non-structural protein 3, a ZIKV non-structural protein 4A, a ZIKV non-structural protein 4B, or a ZIKV non-structural protein 5.

In some embodiments, at least one antigenic polypeptide is a ZIKV capsid protein, a ZIKV premembrane/membrane protein, a ZIKV envelope protein, a ZIKV non-structural protein 1, a ZIKV non-structural protein 2A, a ZIKV non-structural protein 2B, a ZIKV non-structural protein 3, a ZIKV non-structural protein 4A, a ZIKV non-structural protein 4B, or a ZIKV non-structural protein 5.

In some embodiments, the vaccine comprises a RNA polynucleotide having an open reading frame encoding a ZIKV capsid protein, a RNA polynucleotide having an open reading frame encoding a ZIKV premembrane/membrane protein, and a RNA polynucleotide having an open reading frame encoding a ZIKV envelope protein.

In some embodiments, the vaccine comprises a RNA polynucleotide having an open reading frame encoding a ZIKV capsid protein and a RNA polynucleotide having an open reading frame encoding a ZIKV premembrane/membrane protein.

In some embodiments, the vaccine comprises a RNA polynucleotide having an open reading frame encoding a ZIKV capsid protein and a RNA polynucleotide having an open reading frame encoding a ZIKV envelope protein.

In some embodiments, the vaccine comprises a RNA polynucleotide having an open reading frame encoding a ZIKV premembrane/membrane protein and a RNA polynucleotide having an open reading frame encoding a ZIKV envelope protein.

In some embodiments, the vaccine comprises a RNA polynucleotide having an open reading frame encoding a ZIKV capsid protein and at least one RNA polynucleotide having an open reading frame encoding any one or more of a ZIKV non-structural protein 1, 2A, 2B, 3, 4A, 4B or 5.

In some embodiments, the vaccine comprises a RNA polynucleotide having an open reading frame encoding a ZIKV premembrane/membrane protein and at least one RNA polynucleotide having an open reading frame encoding any one or more of a ZIKV non-structural protein 1, 2A, 2B, 3, 4A, 4B or 5.

In some embodiments, the vaccine comprises a RNA polynucleotide having an open reading frame encoding a ZIKV envelope protein and at least one RNA polynucleotide having an open reading frame encoding any one or more of a ZIKV non-structural protein 1, 2A, 2B, 3, 4A, 4B or 5.

In some embodiments, the at least one antigenic polypeptide comprises a combination of any two or more of a ZIKV capsid protein, a ZIKV premembrane/membrane protein, a ZIKV envelope protein, a ZIKV non-structural protein 1, a ZIKV non-structural protein 2A, a ZIKV non-structural protein 2B, a ZIKV non-structural protein 3, a ZIKV non-structural protein 4A, a ZIKV non-structural protein 4B, or a ZIKV non-structural protein 5.

In some embodiments, at least one RNA polynucleotide is encoded by at least one nucleic acid sequence identified by any one of SEQ ID NO: 118-136 (Table 25) and homologs having at least 80% identity with a nucleic acid sequence identified by any one of SEQ ID NO: 118-136 (Table 25). In some embodiments, at least one RNA polynucleotide is encoded by at least one nucleic acid sequence identified by any one of SEQ ID NO: 118-136 (Table 25) and homologs having at least 90% (90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.8% or 99.9%) identity with a nucleic acid sequence identified by any one of SEQ ID NO: 118-136 (Table 25). In some embodiments, at least one RNA polynucleotide is encoded by at least one fragment of a nucleic acid sequence identified by any one of SEQ ID NO: 118-136 (Table 25).

In some embodiments, at least one RNA polynucleotide comprises at least one nucleic acid sequence identified by any one of SEQ ID NO: 137-155 (Table 25) and homologs having at least 80% identity with a nucleic acid sequence identified by any one of SEQ ID NO: 137-155 (Table 25). In some embodiments, at least one RNA polynucleotide comprises at least one nucleic acid sequence identified by any one of SEQ ID NO: 137-155 (Table 25) and homologs having at least 90% (90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.8% or 99.9%) identity with a nucleic acid sequence identified by any one of SEQ ID NO: 137-155 (Table 25). In some embodiments, at least one RNA polynucleotide comprises at least one fragment of a nucleic acid sequence identified by any one of SEQ ID NO: 137-155 (Table 25).

In some embodiments, the at least one RNA polynucleotide encodes at least one antigenic polypeptide having a sequence identified by any one of SEQ ID NO: 156-222 or 469 (Table 26 and 27). In some embodiments, the at least one RNA polynucleotide encodes at least one protein variant having at least 95% identity to an antigenic polypeptide having a sequence identified by any one of SEQ ID NO: 156-222 or 469 (Table 26 and 27). In some embodiments, at least one antigenic polypeptide has an amino acid sequence identified by any one of SEQ ID NO: 156-222 or 469 (Table 26 and 27). In some embodiments, at least one antigenic polypeptide has at least 95% identity to an antigenic polypeptide having a sequence identified by any one of SEQ ID NO: 156-222 or 469 (Table 26 and 27).

Dengue Virus (DENV)

Dengue virus (DENV) is a mosquito-borne (Aedes aegypti/Aedes albopictus) member of the family Flaviviridae (positive-sense, single-stranded RNA virus). Dengue virus is a positive-sense RNA virus of the Flavivirus genus of the Flaviviridae family, which also includes West Nile virus, Yellow Fever Virus, and Japanese Encephalitis virus. It is transmitted to humans through Stegomyia aegypti (formerly Aedes) mosquito vectors and is mainly found in the tropical and semitropical areas of the world, where it is endemic in Asia, the Pacific region, Africa, Latin America, and the Caribbean. The incidence of infections has increased 30-fold over the last 50 years (WHO, Dengue: Guidelines for diagnosis, treatment, prevention, and control (2009)) and Dengue virus is the second most common tropical infectious disease worldwide after malaria.

Severe disease is most commonly observed in secondary, heterologous DENV infections. Antibody-dependent enhancement of infection has been proposed as the primary mechanism of dengue immunopathogenesis. The potential risk of immune enhancement of infection and disease underscores the importance of developing dengue vaccines which produce balanced, long-lasting immunity to at least DENV 1-4, if not all five of the DENV serotypes. While several dengue vaccines are in development, none have been officially licensed and/or approved to date.

In view of the lack of Dengue virus (DENV) vaccines, there is a significant need for a vaccine that would be safe and effective in all patient populations to prevent and/or to treat DENV infection, including those individuals at risk for secondary, heterotypic infections (those with more than one circulating serotype).

Some embodiments of the present disclosure provide Dengue virus (DENV) vaccines that include at least one ribonucleic acid (RNA) polynucleotide (e.g., mRNA polynucleotide) having an open reading frame encoding at least one DENV antigenic polypeptide or an immunogenic fragment thereof (e.g., an immunogenic fragment capable of inducing an immune response to DENV).

The methods of the present disclosure, in some embodiments, enable the production of highly antigenic DENV RNA vaccines, including RNA polynucleotides encoding concatemeric peptide epitopes. The peptide epitopes are designed to be processed intracellularly and presented to the immune system in an efficient manner. The RNA (e.g., mRNA) vaccines described herein are useful for generating a desired immune response by selecting appropriate T or B cell epitopes which are able to be presented more effectively on MHC-I or MHC-II molecules (depending on whether they are T or B-cell epitopes, respectively).

In some embodiments, the at least one RNA polynucleotide encodes a DENV capsid protein or immunogenic fragment or epitope thereof. In some embodiments, the at least one RNA polynucleotide encodes a DENV membrane protein or immunogenic fragment or epitope thereof. In some embodiments, the at least one RNA polynucleotide encodes a DENV precursor-membrane protein or immunogenic fragment or epitope thereof. In some embodiments, the at least one RNA polynucleotide encodes a DENV precursor membrane (prM) and envelope (E) polypeptide (DENV prME) or immunogenic fragment or epitope thereof. In some embodiments, the at least one RNA (e.g., mRNA) polynucleotide encodes a DENV nonstructural protein or immunogenic fragment or epitope thereof, for example a DENV non-structural protein selected from NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5 proteins, or immunogenic fragments or epitopes thereof. In some embodiments, the DENV non-structural protein is NS3.

In some embodiments, the Dengue virus antigen comprises one or more Dengue virus peptide epitopes. In some embodiments, the one or more Dengue virus peptide epitopes is from a DENV envelope protein. In some embodiments, the one or more Dengue virus peptide epitopes is from a DENV capsid protein. In some embodiments, the one or more Dengue virus peptide epitopes is from a DENV membrane protein. In some embodiments, the one or more Dengue virus peptide epitopes is from a DENV pre-membrane protein. In some embodiments, the one or more Dengue virus peptide epitopes is from a sequence comprising DENV precursor membrane (prM) and envelope (E) polypeptide (DENV prME). In some embodiments, the at least one Dengue virus antigen is a DENV2 prME peptide epitope. In some embodiments, the one or more Dengue virus peptide epitopes is from a DENV nonstructural protein.

In any of these embodiments, the at least one RNA (e.g., mRNA) polynucleotide encodes a DENV polypeptide, fragment, or epitope from a DENV serotype selected from DENV-1, DENV-2, DENV-3, DENV-4, and DENV-5. In some embodiments, the one or more Dengue virus peptide epitopes is from a DENV-2 serotype. In some embodiments, the one or more Dengue virus peptide epitopes is from a DENV2 membrane polypeptide. In some embodiments, the one or more Dengue virus peptide epitopes is from a DENV2 envelope polypeptide. In some embodiments, the one or more Dengue virus peptide epitopes is from a DENV2 pre-membrane polypeptide. In some embodiments, the one or more Dengue virus peptide epitopes is from a DENV2 capsid polypeptide. In some embodiments, the one or more Dengue virus peptide epitopes is from a DENV2 non-structural polypeptide. In some embodiments, the one or more Dengue virus peptide epitopes is from a DENV2 pre-membrane polypeptide. In some embodiments, the one or more Dengue virus peptide epitopes is from a DENV2 PrME polypeptide.

In some embodiments, the Dengue virus antigen is a concatemeric Dengue virus antigen comprising two or more Dengue virus peptide epitopes. In some embodiments, the Dengue virus concatemeric antigen comprises between 2-100 Dengue peptide epitopes interspersed by cleavage sensitive sites. In some embodiments, the peptide epitopes are not epitopes of antibody dependent enhancement. In some embodiments, the Dengue virus vaccine's peptide epitopes are T cell epitopes and/or B cell epitopes. In other embodiments, the Dengue virus vaccine's peptide epitopes comprise a combination of T cell epitopes and B cell epitopes. In some embodiments, at least one of the peptide epitopes of the Dengue virus vaccine is a T cell epitope.

In some embodiments, the protease cleavage site of the Dengue virus vaccine comprises the amino acid sequence GFLG (SEQ ID NO: 429), KVSR (SEQ ID NO: 430), TVGLR (SEQ ID NO: 431), PMGLP (SEQ ID NO: 432), or PMGAP (SEQ ID NO: 433).

In some embodiments, the at least one RNA polynucleotide encodes a DENV envelope protein, and one or more concatemeric Dengue virus antigen(s), such as any of the concatemeric antigens described herein. In some embodiments, the at least one RNA polynucleotide encodes a DENV membrane protein, and a concatemeric virus antigen, such as any of the concatemeric antigens described herein. In some embodiments, the at least one RNA polynucleotide encodes a DENV capsid protein and a concatemeric virus antigen, such as any of the concatemeric antigens described herein. In some embodiments, the at least one RNA polynucleotide encodes a DENV nonstructural protein, for example a DENV non-structural protein selected from NS1, NS2A, NS2B, NS3, SN4A, NS4B, and NS5 proteins, and a concatemeric Dengue virus antigen, such as any of the concatemeric antigens described herein. In some embodiments, the DENV non-structural protein is NS3. In some embodiments, the at least one RNA polynucleotide encodes a DENV precursor membrane protein, and one or more concatemeric Dengue virus antigen(s), such as any of the concatemeric antigens described herein. In some embodiments, the at least one RNA polynucleotide encodes a DENV prME polypeptide, and one or more concatemeric Dengue virus antigen(s), such as any of the concatemeric antigens described herein.

In some embodiments, the peptide epitopes comprise at least one MHC class I epitope and at least one MHC class II epitope. In some embodiments, at least 10% of the epitopes are MHC class I epitopes. In some embodiments, at least 20% of the epitopes are MHC class I epitopes. In some embodiments, at least 30% of the epitopes are MHC class I epitopes. In some embodiments, at least 40% of the epitopes are MHC class I epitopes. In some embodiments, at least 50%, 60%, 70%, 80%, 90% or 100% of the epitopes are MHC class I epitopes. In some embodiments, at least 10% of the epitopes are MHC class II epitopes. In some embodiments, at least 20% of the epitopes are MHC class II epitopes. In some embodiments, at least 30% of the epitopes are MHC class II epitopes. In some embodiments, at least 40% of the epitopes are MHC class II epitopes. In some embodiments, at least 50%, 60%, 70%, 80%, 90% or 100% of the epitopes are MHC class II epitopes. In some embodiments, the ratio of MHC class I epitopes to MHC class II epitopes is a ratio selected from about 10%:about 90%; about 20%:about 80%; about 30%:about 70%; about 40%:about 60%; about 50%:about 50%; about 60%:about 40%; about 70%:about 30%; about 80%:about 20%; about 90%:about 10% MHC class 1:MHC class II epitopes. In some embodiments, the ratio of MHC class II epitopes to MHC class I epitopes is a ratio selected from about 10%:about 90%; about 20%:about 80%; about 30%:about 70%; about 40%:about 60%; about 50%:about 50%; about 60%:about 40%; about 70%:about 30%; about 80%:about 20%; about 90%:about 10% MHC class II:MHC class I epitopes. In some embodiments, at least one of the peptide epitopes of the Dengue virus vaccine is a B cell epitope. In some embodiments, the T cell epitope of the Dengue virus vaccine comprises between 8-11 amino acids. In some embodiments, the B cell epitope of the Dengue virus vaccine comprises between 13-17 amino acids.

In any of these embodiments, the concatemeric Dengue virus antigen may comprise two or more Dengue virus peptide epitopes selected from a DENV envelope polypeptide, DENV capsid polypeptide, DENV membrane polypeptide, DENV precursor-membrane polypeptide, DENV nonstructural polypeptide, DENV prME polypeptide, and any combination thereof, and the two or more Dengue virus peptide epitopes may be from any DENV serotype, for example, a DENV serotype selected from DENV-1, DENV-2, DENV-3, DENV-4, DENV-5 and combinations thereof. In some embodiments, the concatemeric Dengue virus antigen comprises two or more Dengue virus peptide epitopes from DENV-2 serotype. In some embodiments, the concatemeric Dengue virus antigen comprises two or more DENV2 prME peptide epitopes, which may be the same or different DENV prME peptide epitopes.

In some embodiments, at least one RNA polynucleotide is encoded by at least one nucleic acid sequence identified by any one of SEQ ID NO: 223-239 (Table 28) and homologs having at least 80% identity with a nucleic acid sequence identified by any one of SEQ ID NO: 223-239 (Table 28). In some embodiments, at least one RNA polynucleotide is encoded by at least one nucleic acid sequence identified by any one of SEQ ID NO: 223-239 (Table 28) and homologs having at least 90% (90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.8% or 99.9%) identity with a nucleic acid sequence identified by any one of SEQ ID NO: 223-239 (Table 28). In some embodiments, at least one RNA polynucleotide is encoded by at least one fragment of a nucleic acid sequence identified by any one of SEQ ID NO: 223-239 (Table 28).

In some embodiments, at least one RNA polynucleotide comprises at least one nucleic acid sequence identified by any one of SEQ ID NO: 240-256 (Table 28) and homologs having at least 80% identity with a nucleic acid sequence identified by any one of SEQ ID NO: 240-256 (Table 28). In some embodiments, at least one RNA polynucleotide comprises at least one nucleic acid sequence identified by any one of SEQ ID NO: 240-256 (Table 28) and homologs having at least 90% (90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.8% or 99.9%) identity with a nucleic acid sequence identified by any one of SEQ ID NO: 240-256 (Table 28). In some embodiments, at least one RNA polynucleotide comprises at least one fragment of a nucleic acid sequence identified by any one of SEQ ID NO: 240-256 (Table 28).

In some embodiments, the at least one RNA polynucleotide encodes at least one antigenic polypeptide having a sequence identified by any one of SEQ ID NO: 259-291 (Table 29 and 42). In some embodiments, the at least one RNA polynucleotide encodes at least one protein variant having at least 95% identity to an antigenic polypeptide having a sequence identified by any one of SEQ ID NO: 259-291 (Table 29 and 42). In some embodiments, at least one antigenic polypeptide has an amino acid sequence identified by any one of SEQ ID NO: 259-291 (Table 29 and 42). In some embodiments, at least one antigenic polypeptide has at least 95% identity to an antigenic polypeptide having a sequence identified by any one of SEQ ID NO: 259-291 (Table 29 and 42).

Chikungunya Virus (CHIKV)

Some embodiments of the present disclosure provide Chikungunya virus (CHIKV) vaccines that include at least one ribonucleic acid (RNA) polynucleotide (e.g., mRNA polynucleotide) having an open reading frame encoding at least one CHIKV antigenic polypeptide or an immunogenic fragment thereof (e.g., an immunogenic fragment capable of inducing an immune response to CHIKV).

Chikungunya virus (CHIKV) is a mosquito-borne virus belonging to the Alphavirus genus of the Togaviridae family that was first isolated in 1953 in Tanzania, where the virus was endemic. Outbreaks occur repeatedly in west, central, and southern Africa and have caused several human epidemics in those areas since that time. The virus is passed to humans by two species of mosquito of the genus Aedes: A. albopictus and A. aegypti. There are several Chikungunya genotypes: Indian Ocean, East/Central/South African (ECSA), Asian, West African, and Brazilian.

The CHIKV antigenic polypeptide may be a Chikungunya structural protein or an antigenic fragment or epitope thereof. In some embodiments, the antigenic polypeptide is a CHIKV structural protein or an antigenic fragment thereof. For example, a CHIKV structural protein may be an envelope protein (E), a 6K protein, or a capsid (C) protein. In some embodiments, the CHIKV structural protein is an envelope protein selected from E1, E2, and E3. In some embodiments, the CHIKV structural protein is E1 or E2. In some embodiments, the CHIKV structural protein is a capsid protein. In some embodiments, the antigenic polypeptide is a fragment or epitope of a CHIKV structural protein.

In some embodiments, the antigenic polypeptide comprises two or more CHIKV structural proteins. In some embodiments, the two or more CHIKV structural proteins are envelope proteins. In some embodiments, the two or more CHIKV structural proteins are E1 and E2. In some embodiments, the two or more CHIKV structural proteins are E1 and E3. In some embodiments, the two or more CHIKV structural proteins are E2 and E3. In some embodiments, the two or more CHIKV structural proteins are E1, E2, and E3. In some embodiments, the two or more CHIKV structural proteins are envelope and capsid proteins. In some embodiments, the two or more CHIKV structural proteins are E1 and C. In some embodiments, the two or more CHIKV structural proteins are E2 and C. In some embodiments, the two or more CHIKV structural proteins are E3 and C. In some embodiments, the two or more CHIKV structural proteins are E1, E2, and C. In some embodiments, the two or more CHIKV structural proteins are E1, E3, and C. In some embodiments, the two or more CHIKV structural proteins are E2, E3, and C. In some embodiments, the two or more CHIKV structural proteins are E1, E2, E3, and C. In some embodiments, the two or more CHIKV structural proteins are E1, 6K, and E2. In some embodiments, the two or more CHIKV structural proteins are E2, 6K, and E3. In some embodiments, the two or more CHIKV structural proteins are E1, 6K, and E3. In some embodiments, the two or more CHIKV structural proteins are E1, E2, E3, 6K, and C. In some embodiments, the antigenic polypeptide comprises the CHIKV structural polyprotein comprising C, E3, E2, 6K, and E1. In some embodiments, the antigenic polypeptide is a fragment or epitope of two or more CHIKV structural proteins or a fragment or epitope of the polyprotein.

In some embodiments, at least one RNA polynucleotide is encoded by at least one nucleic acid sequence identified by any one of SEQ ID NO: 376-388 (Table 47) and homologs having at least 80% identity with a nucleic acid sequence identified by any one of SEQ ID NO: 376-388 (Table 47). In some embodiments, at least one RNA polynucleotide is encoded by at least one nucleic acid sequence identified by any one of SEQ ID NO: 376-388 (Table 47) and homologs having at least 90% (90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.8% or 99.9%) identity with a nucleic acid sequence identified by any one of SEQ ID NO: 376-388 (Table 47). In some embodiments, at least one RNA polynucleotide is encoded by at least one fragment of a nucleic acid sequence identified by any one of SEQ ID NO: 376-388 (Table 47).

In some embodiments, at least one RNA polynucleotide comprises at least one nucleic acid sequence identified by any one of SEQ ID NO: 389-401 (Table 47) and homologs having at least 80% identity with a nucleic acid sequence identified by any one of SEQ ID NO: 389-401 (Table 47). In some embodiments, at least one RNA polynucleotide comprises at least one nucleic acid sequence identified by any one of SEQ ID NO: 389-401 (Table 47) and homologs having at least 90% (90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.8% or 99.9%) identity with a nucleic acid sequence identified by any one of SEQ ID NO: 389-401 (Table 47). In some embodiments, at least one RNA polynucleotide comprises at least one fragment of a nucleic acid sequence identified by any one of SEQ ID NO: 389-401 (Table 47).

In some embodiments, the at least one RNA polynucleotide encodes at least one antigenic polypeptide having a sequence identified by any one of SEQ ID NO: 402-413 (Table 48). In some embodiments, the at least one RNA polynucleotide encodes at least one protein variant having at least 95% identity to an antigenic polypeptide having a sequence identified by any one of SEQ ID NO: 402-413 (Table 48). In some embodiments, at least one antigenic polypeptide has an amino acid sequence identified by any one of SEQ ID NO: 402-413 (Table 48). In some embodiments, at least one antigenic polypeptide has at least 95% identity to an antigenic polypeptide having a sequence identified by any one of SEQ ID NO: 402-413 (Table 48).

In some embodiments, an open reading frame of a RNA (e.g., mRNA) vaccine is codon-optimized. In some embodiments, at least one RNA polynucleotide encodes at least one antigenic polypeptide comprising an amino acid sequence identified by any one of SEQ ID NO: 13-17, 22-29, 44-47, 52-54, 59-64, 97-117, 156-222, 469, 259-291 or 402-413 and is codon optimized mRNA.

In some embodiments, a RNA (e.g., mRNA) vaccine further comprising an adjuvant.

Tables 3, 6, 11, 14, 17, 27, and 42 provide National Center for Biotechnology Information (NCBI) accession numbers of interest. It should be understood that the phrase “an amino acid sequence of Tables 3, 6, 11, 14, 17, 27, and 42” refers to an amino acid sequence identified by one or more NCBI accession numbers listed in Tables 3, 6, 11, 14, 17, 27, and 42. Each of the amino acid sequences, and variants having greater than 95% identity or greater than 98% identity to each of the amino acid sequences encompassed by the accession numbers of Tables 3, 6, 11, 14, 17, 27, and 42 are included within the constructs (polynucleotides/polypeptides) of the present disclosure.

In some embodiments, at least one mRNA polynucleotide is encoded by a nucleic acid comprising a sequence identified by any one of SEQ ID NO: 1-6, 18, 19, 30-34, 48, 49, 55, 56, 65-80, 118-136, 223-239 or 376-388 and having less than 80% identity to wild-type mRNA sequence. In some embodiments, at least one mRNA polynucleotide is encoded by a nucleic acid comprising a sequence identified by any one of SEQ ID NO: 1-6, 18, 19, 30-34, 48, 49, 55, 56, 65-80, 118-136, 223-239 or 376-388 and having less than 75%, 85% or 95% identity to a wild-type mRNA sequence. In some embodiments, at least one mRNA polynucleotide is encoded by nucleic acid comprising a sequence identified by any one of SEQ ID NO: 1-6, 18, 19, 30-34, 48, 49, 55, 56, 65-80, 118-136, 223-239 or 376-388 and having less than 50-80%, 60-80%, 40-80%, 30-80%, 70-80%, 75-80% or 78-80% identity to wild-type mRNA sequence. In some embodiments, at least one mRNA polynucleotide is encoded by a nucleic acid comprising a sequence identified by any one of SEQ ID NO: 1-6, 18, 19, 30-34, 48, 49, 55, 56, 65-80, 118-136, 223-239 or 376-388 and having less than 40-85%, 50-85%, 60-85%, 30-85%, 70-85%, 75-85% or 80-85% identity to wild-type mRNA sequence. In some embodiments, at least one mRNA polynucleotide is encoded by a nucleic acid comprising a sequence identified by any one of SEQ ID NO: 1-6, 18, 19, 30-34, 48, 49, 55, 56, 65-80, 118-136, 223-239 or 376-388 and having less than 40-90%, 50-90%, 60-90%, 30-90%, 70-90%, 75-90%, 80-90%, or 85-90% identity to wild-type mRNA sequence.

In some embodiments, at least one mRNA polynucleotide comprises a nucleic acid comprising a sequence identified by any one of SEQ ID NO: 7-12, 20-21, 35-39, 50-51, 57-58, 81-96, 137-155, 240-256, or 389-401 (with or without a signal sequence, 5′ UTR, 3′ UTR, and/or polyA tail) and having less than 80% identity to wild-type mRNA sequence. In some embodiments, at least one mRNA polynucleotide comprises a nucleic acid comprising a sequence identified by any one of SEQ ID NO: 7-12, 20-21, 35-39, 50-51, 57-58, 81-96, 137-155, 240-256, or 389-401 (with or without a signal sequence, 5′ UTR, 3′ UTR, and/or polyA tail) and having less than 75%, 85% or 95% identity to a wild-type mRNA sequence. In some embodiments, at least one mRNA polynucleotide comprises nucleic acid comprising a sequence identified by any one of SEQ ID NO: 7-12, 20-21, 35-39, 50-51, 57-58, 81-96, 137-155, 240-256, or 389-401 (with or without a signal sequence, 5′ UTR, 3′ UTR, and/or polyA tail) and having less than 50-80%, 60-80%, 40-80%, 30-80%, 70-80%, 75-80% or 78-80% identity to wild-type mRNA sequence. In some embodiments, at least one mRNA polynucleotide comprises a nucleic acid comprising a sequence identified by any one of SEQ ID NO: 7-12, 20-21, 35-39, 50-51, 57-58, 81-96, 137-155, 240-256, or 389-401 (with or without a signal sequence, 5′ UTR, 3′ UTR, and/or polyA tail) and having less than 40-85%, 50-85%, 60-85%, 30-85%, 70-85%, 75-85% or 80-85% identity to wild-type mRNA sequence. In some embodiments, at least one mRNA polynucleotide comprises a nucleic acid comprising a sequence identified by any one of SEQ ID NO: 7-12, 20-21, 35-39, 50-51, 57-58, 81-96, 137-155, 240-256, or 389-401 (with or without a signal sequence, 5′ UTR, 3′ UTR, and/or polyA tail) and having less than 40-90%, 50-90%, 60-90%, 30-90%, 70-90%, 75-90%, 80-90%, or 85-90% identity to wild-type mRNA sequence.

In some embodiments, at least one RNA polynucleotide encodes at least one antigenic polypeptide comprising an amino acid sequence identified by any one of SEQ ID NO: 13-17, 22-29, 44-47, 52-54, 59-64, 97-117, 156-222, 469, 259-291 or 402-413 and having at least 80% (e.g., 85%, 90%, 95%, 98%, 99%) identity to wild-type mRNA sequence, but does not include wild-type mRNA sequence.

In some embodiments, at least one RNA polynucleotide encodes at least one antigenic polypeptide comprising an amino acid sequence identified by any one of SEQ ID NO: 13-17, 22-29, 44-47, 52-54, 59-64, 97-117, 156-222, 469, 259-291 or 402-413 and has less than 95%, 90%, 85%, 80% or 75% identity to wild-type mRNA sequence. In some embodiments, at least one RNA polynucleotide encodes at least one antigenic polypeptide comprising an amino acid sequence identified by any one of SEQ ID NO: 13-17, 22-29, 44-47, 52-54, 59-64, 97-117, 156-222, 469, 259-291 or 402-413 and has 30-80%, 40-80%, 50-80%, 60-80%, 70-80%, 75-80% or 78-80%, 30-85%, 40-85%, 50-85%, 60-85%, 70-85%, 75-85% or 78-85%, 30-90%, 40-90%, 50-90%, 60-90%, 70-90%, 75-90%, 80-90% or 85-90% identity to wild-type mRNA sequence.

In some embodiments, at least one RNA polynucleotide encodes at least one antigenic polypeptide having at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to an amino acid sequence identified by any one of SEQ ID NO: 13-17, 22-29, 44-47, 52-54, 59-64, 97-117, 156-222, 469, 259-291 or 402-413. In some embodiments, at least one RNA polynucleotide encodes at least one antigenic polypeptide having 95-99% identity to an amino acid sequence identified by any one of SEQ ID NO: 13-17, 22-29, 44-47, 52-54, 59-64, 97-117, 156-222, 469, 259-291 or 402-413.

In some embodiments, at least one RNA polynucleotide encodes at least one antigenic polypeptide having at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to amino acid sequence identified by any one of SEQ ID NO: 13-17, 22-29, 44-47, 52-54, 59-64, 97-117, 156-222, 469, 259-291 or 402-413 and having membrane fusion activity. In some embodiments, at least one RNA polynucleotide encodes at least one antigenic polypeptide having 95-99% identity to amino acid sequence identified by any one of SEQ ID NO: 13-17, 22-29, 44-47, 52-54, 59-64, 97-117, 156-222, 469, 259-291 or 402-413 and having membrane fusion activity.

In some embodiments, at least one RNA polynucleotide encodes at least one antigenic polypeptide (e.g., at least one Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide) that attaches to cell receptors.

In some embodiments, at least one RNA polynucleotide encodes at least one antigenic polypeptide (e.g., at least one Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide) antigenic polypeptide) that causes fusion of viral and cellular membranes.

In some embodiments, at least one RNA polynucleotide encodes at least one antigenic polypeptide (e.g., at least one Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide) that is responsible for binding of the virus to a cell being infected.

Some embodiments of the present disclosure provide a vaccine that includes at least one ribonucleic acid (RNA) (e.g., mRNA) polynucleotide having an open reading frame encoding at least one antigenic polypeptide (e.g., at least one Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide), at least one 5′ terminal cap and at least one chemical modification, formulated within a lipid nanoparticle.

In some embodiments, a 5′ terminal cap is 7mG(5′)ppp(5′)NlmpNp.

In some embodiments, at least one chemical modification is selected from pseudouridine, N1-methylpseudouridine, N1-ethylpseudouridine, 2-thiouridine, 4′-thiouridine, 5-methylcytosine, 5-methyluridine, 2-thio-1-methyl-1-deaza-pseudouridine, 2-thio-1-methyl-pseudouridine, 2-thio-5-aza-uridine, 2-thio-dihydropseudouridine, 2-thio-dihydrouridine, 2-thio-pseudouridine, 4-methoxy-2-thio-pseudouridine, 4-methoxy-pseudouridine, 4-thio-1-methyl-pseudouridine, 4-thio-pseudouridine, 5-aza-uridine, dihydropseudouridine, 5-methoxyuridine and 2′-O-methyl uridine. In some embodiments, the chemical modification is in the 5-position of the uracil. In some embodiments, the chemical modification is a N1-methylpseudouridine or a N1-ethylpseudouridine. In some embodiments, a lipid nanoparticle comprises a cationic lipid, a PEG-modified lipid, a sterol and a non-cationic lipid. In some embodiments, a cationic lipid is an ionizable cationic lipid and the non-cationic lipid is a neutral lipid, and the sterol is a cholesterol. In some embodiments, a cationic lipid is selected from the group consisting of 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), (12Z,15Z)—N,N-dimethyl-2-nonylhenicosa-12,15-dien-1-amine (L608), and N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]heptadecan-8-amine (L530).

In some embodiments, the lipid is

In some embodiments, the lipid is

In some embodiments, a lipid nanoparticle comprises compounds of Formula (I) and/or Formula (II), discussed below.

In some embodiments, a lipid nanoparticle comprises Compounds 3, 18, 20, 25, 26, 29, 30, 60, 108-112, or 122, as discussed below.

Some embodiments of the present disclosure provide a vaccine that includes at least one RNA (e.g., mRNA) polynucleotide having an open reading frame encoding at least one antigenic polypeptide (e.g., at least one Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide), wherein at least 80% (e.g., 85%, 90%, 95%, 98%, 99%) of the uracil in the open reading frame have a chemical modification, optionally wherein the vaccine is formulated in a lipid nanoparticle (e.g., a lipid nanoparticle comprises a cationic lipid, a PEG-modified lipid, a sterol and a non-cationic lipid).

In some embodiments, 100% of the uracil in the open reading frame have a chemical modification. In some embodiments, a chemical modification is in the 5-position of the uracil. In some embodiments, a chemical modification is a N1-methyl pseudouridine. In some embodiments, 100% of the uracil in the open reading frame have a N1-methyl pseudouridine in the 5-position of the uracil.

In some embodiments, an open reading frame of a RNA (e.g., mRNA) polynucleotide encodes at least two antigenic polypeptides (e.g., at least one Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide). In some embodiments, the open reading frame encodes at least five or at least ten antigenic polypeptides. In some embodiments, the open reading frame encodes at least 100 antigenic polypeptides. In some embodiments, the open reading frame encodes 2-100 antigenic polypeptides.

In some embodiments, a vaccine comprises at least two RNA (e.g., mRNA) polynucleotides, each having an open reading frame encoding at least one antigenic polypeptide (e.g., at least one Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide). In some embodiments, the vaccine comprises at least five or at least ten RNA (e.g., mRNA) polynucleotides, each having an open reading frame encoding at least one antigenic polypeptide or an immunogenic fragment thereof. In some embodiments, the vaccine comprises at least 100 RNA (e.g., mRNA) polynucleotides, each having an open reading frame encoding at least one antigenic polypeptide. In some embodiments, the vaccine comprises 2-100 RNA (e.g., mRNA) polynucleotides, each having an open reading frame encoding at least one antigenic polypeptide.

In some embodiments, at least one antigenic polypeptide (e.g., at least one Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide) is fused to a signal peptide. In some embodiments, the signal peptide is selected from: a HuIgGk signal peptide (METPAQLLFLLLLWLPDTTG; SEQ ID NO: 423); IgE heavy chain epsilon-1 signal peptide (MDWTWILFLVAAATRVHS; SEQ ID NO: 424); Japanese encephalitis PRM signal sequence (MLGSNSGQRVVFTILLLLVAPAYS; SEQ ID NO: 425), VSINVg protein signal sequence (MKCLLYLAFLFIGVNCA; SEQ ID NO: 426) and Japanese encephalitis JEV signal sequence (MWLVSLAIVTACAGA; SEQ ID NO: 427).

In some embodiments, the signal peptide is fused to the N-terminus of at least one antigenic polypeptide. In some embodiments, a signal peptide is fused to the C-terminus of at least one antigenic polypeptide.

In some embodiments, at least one antigenic polypeptide (e.g., at least one Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide) comprises a mutated N-linked glycosylation site.

Also provided herein is a RNA (e.g., mRNA) vaccine of any one of the foregoing paragraphs (e.g., at least one Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide), formulated in a nanoparticle (e.g., a lipid nanoparticle).

In some embodiments, the nanoparticle has a mean diameter of 50-200 nm. In some embodiments, the nanoparticle is a lipid nanoparticle.

In some embodiments, a lipid nanoparticle comprises compounds of Formula (I) and/or Formula (II), discussed below.

In some embodiments, a tropical disease RNA (e.g., mRNA) vaccine is formulated in a lipid nanoparticle that comprises a compound selected from Compounds 3, 18, 20, 25, 26, 29, 30, 60, 108-112 and 122, described below.

In some embodiments, the nanoparticle has a polydispersity value of less than 0.4 (e.g., less than 0.3, 0.2 or 0.1).

In some embodiments, the nanoparticle has a net neutral charge at a neutral pH value.

In some embodiments, the RNA (e.g., mRNA) vaccine is multivalent.

Some embodiments of the present disclosure provide methods of inducing an antigen specific immune response in a subject, comprising administering to the subject any of the RNA (e.g., mRNA) vaccine as provided herein in an amount effective to produce an antigen-specific immune response. In some embodiments, the RNA (e.g., mRNA) vaccine is a Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV vaccine. In some embodiments, the RNA (e.g., mRNA) vaccine is a combination vaccine comprising a combination of Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale) vaccine, JEV vaccine, WNV vaccine, EEEV vaccine, SINV vaccine, CHIKV vaccine, DENV vaccine, ZIKV vaccine and/or YFV vaccine.

In some embodiments, an antigen-specific immune response comprises a T cell response or a B cell response.

In some embodiments, a method of producing an antigen-specific immune response comprises administering to a subject a single dose (no booster dose) of a RNA (e.g., mRNA) vaccine of the present disclosure. In some embodiments, the RNA (e.g., mRNA) vaccine is a Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale) vaccine, JEV vaccine, WNV vaccine, EEEV vaccine, SINV vaccine, CHIKV vaccine, DENV vaccine, ZIKV vaccine and/or YFV vaccine. In some embodiments, the RNA (e.g., mRNA) vaccine is a combination vaccine comprising a combination of any two or more of the foregoing vaccines.

In some embodiments, a method further comprises administering to the subject a second (booster) dose of a RNA (e.g., mRNA) vaccine. Additional doses of a RNA (e.g., mRNA) vaccine may be administered.

In some embodiments, the subjects exhibit a seroconversion rate of at least 80% (e.g., at least 85%, at least 90%, or at least 95%) following the first dose or the second (booster) dose of the vaccine. Seroconversion is the time period during which a specific antibody develops and becomes detectable in the blood. After seroconversion has occurred, a virus can be detected in blood tests for the antibody. During an infection or immunization, antigens enter the blood, and the immune system begins to produce antibodies in response. Before seroconversion, the antigen itself may or may not be detectable, but antibodies are considered absent. During seroconversion, antibodies are present but not yet detectable. Any time after seroconversion, the antibodies can be detected in the blood, indicating a prior or current infection.

In some embodiments, a RNA (e.g., mRNA) vaccine is administered to a subject by intradermal, intramuscular injection, or by intranasal administration.

Some embodiments of the present disclosure provide methods of inducing an antigen specific immune response in a subject, including administering to a subject a RNA (e.g., mRNA) vaccine in an effective amount to produce an antigen specific immune response in a subject. Antigen-specific immune responses in a subject may be determined, in some embodiments, by assaying for antibody titer (for titer of an antibody that binds to a Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide) following administration to the subject of any of the RNA (e.g., mRNA) vaccines of the present disclosure. In some embodiments, the anti-antigenic polypeptide antibody titer produced in the subject is increased by at least 1 log relative to a control. In some embodiments, the anti-antigenic polypeptide antibody titer produced in the subject is increased by 1-3 log relative to a control.

In some embodiments, the anti-antigenic polypeptide antibody titer produced in a subject is increased at least 2 times relative to a control. In some embodiments, the anti-antigenic polypeptide antibody titer produced in the subject is increased at least 5 times relative to a control. In some embodiments, the anti-antigenic polypeptide antibody titer produced in the subject is increased at least 10 times relative to a control. In some embodiments, the anti-antigenic polypeptide antibody titer produced in the subject is increased 2-10 times relative to a control.

In some embodiments, the control is an anti-antigenic polypeptide antibody titer produced in a subject who has not been administered a RNA (e.g., mRNA) vaccine of the present disclosure. In some embodiments, the control is an anti-antigenic polypeptide antibody titer produced in a subject who has been administered a live attenuated or inactivated Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV vaccine (see, e.g., Ren J. et al. J of Gen. Virol. 2015; 96: 1515-1520), or wherein the control is an anti-antigenic polypeptide antibody titer produced in a subject who has been administered a recombinant or purified Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV protein vaccine. In some embodiments, the control is an anti-antigenic polypeptide antibody titer produced in a subject who has been administered a Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV virus-like particle (VLP) vaccine (see, e.g., Cox R G et al., J Virol. 2014 June; 88(11): 6368-6379).

A RNA (e.g., mRNA) vaccine of the present disclosure is administered to a subject in an effective amount (an amount effective to induce an immune response). In some embodiments, the effective amount is a dose equivalent to an at least 2-fold, at least 4-fold, at least 10-fold, at least 100-fold, at least 1000-fold reduction in the standard of care dose of a recombinant Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV protein vaccine, wherein the anti-antigenic polypeptide antibody titer produced in the subject is equivalent to an anti-antigenic polypeptide antibody titer produced in a control subject administered the standard of care dose of a recombinant Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV protein vaccine, a purified Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV protein vaccine, a live attenuated Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV vaccine, an inactivated Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV vaccine, or a Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV VLP vaccine. In some embodiments, the effective amount is a dose equivalent to 2-1000-fold reduction in the standard of care dose of a recombinant Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV protein vaccine, wherein the anti-antigenic polypeptide antibody titer produced in the subject is equivalent to an anti-antigenic polypeptide antibody titer produced in a control subject administered the standard of care dose of a recombinant Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV protein vaccine, a purified Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV protein vaccine, a live attenuated Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV vaccine, an inactivated Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV vaccine, or a Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV VLP vaccine.

In some embodiments, the control is an anti-antigenic polypeptide antibody titer produced in a subject who has been administered a virus-like particle (VLP) vaccine comprising structural proteins of Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV.

In some embodiments, the RNA (e.g., mRNA) vaccine is formulated in an effective amount to produce an antigen specific immune response in a subject.

In some embodiments, the effective amount is a total dose of 25 μg to 1000 μg, or 50 μg to 1000 μg. In some embodiments, the effective amount is a total dose of 100 μg. In some embodiments, the effective amount is a dose of 25 μg administered to the subject a total of two times. In some embodiments, the effective amount is a dose of 100 μg administered to the subject a total of two times. In some embodiments, the effective amount is a dose of 400 μg administered to the subject a total of two times. In some embodiments, the effective amount is a dose of 500 μg administered to the subject a total of two times.

In some embodiments, the efficacy (or effectiveness) of a RNA (e.g., mRNA) vaccine is greater than 60%. In some embodiments, the RNA (e.g., mRNA) polynucleotide of the vaccine is at least one of Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide.

Vaccine efficacy may be assessed using standard analyses (see, e.g., Weinberg et al., J Infect Dis. 2010 Jun. 1; 201(11):1607-10). For example, vaccine efficacy may be measured by double-blind, randomized, clinical controlled trials. Vaccine efficacy may be expressed as a proportionate reduction in disease attack rate (AR) between the unvaccinated (ARU) and vaccinated (ARV) study cohorts and can be calculated from the relative risk (RR) of disease among the vaccinated group with use of the following formulas:

Efficacy=(ARU−ARV)/ARU×100; and

Efficacy=(1−RR)×100.

Likewise, vaccine effectiveness may be assessed using standard analyses (see, e.g., Weinberg et al., J Infect Dis. 2010 Jun. 1; 201(11):1607-10). Vaccine effectiveness is an assessment of how a vaccine (which may have already proven to have high vaccine efficacy) reduces disease in a population. This measure can assess the net balance of benefits and adverse effects of a vaccination program, not just the vaccine itself, under natural field conditions rather than in a controlled clinical trial. Vaccine effectiveness is proportional to vaccine efficacy (potency) but is also affected by how well target groups in the population are immunized, as well as by other non-vaccine-related factors that influence the ‘real-world’ outcomes of hospitalizations, ambulatory visits, or costs. For example, a retrospective case control analysis may be used, in which the rates of vaccination among a set of infected cases and appropriate controls are compared. Vaccine effectiveness may be expressed as a rate difference, with use of the odds ratio (OR) for developing infection despite vaccination:

Effectiveness=(1−OR)×100.

In some embodiments, the efficacy (or effectiveness) of a RNA (e.g., mRNA) vaccine is at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, or at least 90%.

In some embodiments, the vaccine immunizes the subject against Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV for up to 2 years. In some embodiments, the vaccine immunizes the subject against Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV for more than 2 years, more than 3 years, more than 4 years, or for 5-10 years.

In some embodiments, the subject is about 5 years old or younger. For example, the subject may be between the ages of about 1 year and about 5 years (e.g., about 1, 2, 3, 4 or 5 years), or between the ages of about 6 months and about 1 year (e.g., about 6, 7, 8, 9, 10, 11 or 12 months). In some embodiments, the subject is about 12 months or younger (e.g., 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2 months or 1 month). In some embodiments, the subject is about 6 months or younger.

In some embodiments, the subject was born full term (e.g., about 37-42 weeks). In some embodiments, the subject was born prematurely, for example, at about 36 weeks of gestation or earlier (e.g., about 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26 or 25 weeks). For example, the subject may have been born at about 32 weeks of gestation or earlier. In some embodiments, the subject was born prematurely between about 32 weeks and about 36 weeks of gestation. In such subjects, a RNA (e.g., mRNA) vaccine may be administered later in life, for example, at the age of about 6 months to about 5 years, or older.

In some embodiments, the subject is an adult between the ages of about 20 years and about 50 years (e.g., about 20, 25, 30, 35, 40, 45 or 50 years old).

In some embodiments, the subject is an elderly subject about 60 years old, about 70 years old, or older (e.g., about 60, 65, 70, 75, 80, 85 or 90 years old).

In some embodiments, the subject has been exposed to Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV (e.g., C. trachomatis); the subject is infected with Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV (e.g., C. trachomatis); or subject is at risk of infection by Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV (e.g., C. trachomatis).

In some embodiments, the subject is immunocompromised (has an impaired immune system, e.g., has an immune disorder or autoimmune disorder).

In some embodiments the nucleic acid vaccines described herein are chemically modified. In other embodiments the nucleic acid vaccines are unmodified.

Yet other aspects provide compositions for and methods of vaccinating a subject comprising administering to the subject a nucleic acid vaccine comprising one or more RNA polynucleotides having an open reading frame encoding a first virus antigenic polypeptide, wherein the RNA polynucleotide does not include a stabilization element, and wherein an adjuvant is not coformulated or co-administered with the vaccine.

In other aspects the invention is a composition for or method of vaccinating a subject comprising administering to the subject a nucleic acid vaccine comprising one or more RNA polynucleotides having an open reading frame encoding a first antigenic polypeptide wherein a dosage of between 10 μg/kg and 400 μg/kg of the nucleic acid vaccine is administered to the subject. In some embodiments the dosage of the RNA polynucleotide is 1-5 μg, 5-10 μg, 10-15 μg, 15-20 μg, 10-25 μg, 20-25 μg, 20-50 μg, 30-50 μg, 40-50 μg, 40-60 μg, 60-80 μg, 60-100 μg, 50-100 μg, 80-120 μg, 40-120 μg, 40-150 μg, 50-150 μg, 50-200 μg, 80-200 μg, 100-200 μg, 120-250 μg, 150-250 μg, 180-280 μg, 200-300 μg, 50-300 μg, 80-300 μg, 100-300 μg, 40-300 μg, 50-350 μg, 100-350 μg, 200-350 μg, 300-350 μg, 320-400 μg, 40-380 μg, 40-100 μg, 100-400 μg, 200-400 μg, or 300-400 μg per dose. In some embodiments, the nucleic acid vaccine is administered to the subject by intradermal or intramuscular injection. In some embodiments, the nucleic acid vaccine is administered to the subject on day zero. In some embodiments, a second dose of the nucleic acid vaccine is administered to the subject on day twenty one.

In some embodiments, a dosage of 25 micrograms of the RNA polynucleotide is included in the nucleic acid vaccine administered to the subject. In some embodiments, a dosage of 100 micrograms of the RNA polynucleotide is included in the nucleic acid vaccine administered to the subject. In some embodiments, a dosage of 50 micrograms of the RNA polynucleotide is included in the nucleic acid vaccine administered to the subject. In some embodiments, a dosage of 75 micrograms of the RNA polynucleotide is included in the nucleic acid vaccine administered to the subject. In some embodiments, a dosage of 150 micrograms of the RNA polynucleotide is included in the nucleic acid vaccine administered to the subject. In some embodiments, a dosage of 400 micrograms of the RNA polynucleotide is included in the nucleic acid vaccine administered to the subject. In some embodiments, a dosage of 200 micrograms of the RNA polynucleotide is included in the nucleic acid vaccine administered to the subject. In some embodiments, the RNA polynucleotide accumulates at a 100 fold higher level in the local lymph node in comparison with the distal lymph node. In other embodiments the nucleic acid vaccine is chemically modified and in other embodiments the nucleic acid vaccine is not chemically modified.

Aspects of the invention provide a nucleic acid vaccine comprising one or more RNA polynucleotides having an open reading frame encoding a first antigenic polypeptide, wherein the RNA polynucleotide does not include a stabilization element, and a pharmaceutically acceptable carrier or excipient, wherein an adjuvant is not included in the vaccine. In some embodiments, the stabilization element is a histone stem-loop. In some embodiments, the stabilization element is a nucleic acid sequence having increased GC content relative to wild type sequence.

Aspects of the invention provide nucleic acid vaccines comprising one or more RNA polynucleotides having an open reading frame encoding a first antigenic polypeptide, wherein the RNA polynucleotide is present in the formulation for in vivo administration to a host, which confers an antibody titer superior to the criterion for seroprotection for the first antigen for an acceptable percentage of human subjects. In some embodiments, the antibody titer produced by the mRNA vaccines of the invention is a neutralizing antibody titer. In some embodiments the neutralizing antibody titer is greater than a protein vaccine. In other embodiments the neutralizing antibody titer produced by the mRNA vaccines of the invention is greater than an adjuvanted protein vaccine. In yet other embodiments the neutralizing antibody titer produced by the mRNA vaccines of the invention is 1,000-10,000, 1,200-10,000, 1,400-10,000, 1,500-10,000, 1,000-5,000, 1,000-4,000, 1,800-10,000, 2000-10,000, 2,000-5,000, 2,000-3,000, 2,000-4,000, 3,000-5,000, 3,000-4,000, or 2,000-2,500. A neutralization titer is typically expressed as the highest serum dilution required to achieve a 50% reduction in the number of plaques.

Also provided are nucleic acid vaccines comprising one or more RNA polynucleotides having an open reading frame encoding a first antigenic polypeptide, wherein the RNA polynucleotide is present in a formulation for in vivo administration to a host for eliciting a longer lasting high antibody titer than an antibody titer elicited by an mRNA vaccine having a stabilizing element or formulated with an adjuvant and encoding the first antigenic polypeptide. In some embodiments, the RNA polynucleotide is formulated to produce neutralizing antibodies within one week of a single administration. In some embodiments, the adjuvant is selected from a cationic peptide and an immunostimulatory nucleic acid. In some embodiments, the cationic peptide is protamine.

Aspects provide nucleic acid vaccines comprising one or more RNA polynucleotides having an open reading frame comprising at least one chemical modification or optionally no modified nucleotides, the open reading frame encoding a first antigenic polypeptide, wherein the RNA polynucleotide is present in the formulation for in vivo administration to a host such that the level of antigen expression in the host significantly exceeds a level of antigen expression produced by an mRNA vaccine having a stabilizing element or formulated with an adjuvant and encoding the first antigenic polypeptide.

Other aspects provide nucleic acid vaccines comprising one or more RNA polynucleotides having an open reading frame comprising at least one chemical modification or optionally no modified nucleotides, the open reading frame encoding a first antigenic polypeptide, wherein the vaccine has at least 10 fold less RNA polynucleotide than is required for an unmodified mRNA vaccine to produce an equivalent antibody titer. In some embodiments, the RNA polynucleotide is present in a dosage of 25-100 micrograms.

Aspects of the invention also provide a unit of use vaccine, comprising between 10 ug and 400 ug of one or more RNA polynucleotides having an open reading frame comprising at least one chemical modification or optionally no modified nucleotides, the open reading frame encoding a first antigenic polypeptide, and a pharmaceutically acceptable carrier or excipient, formulated for delivery to a human subject. In some embodiments, the vaccine further comprises a cationic lipid nanoparticle.

Aspects of the invention provide methods of creating, maintaining or restoring antigenic memory to a virus strain in an individual or population of individuals comprising administering to said individual or population an antigenic memory booster nucleic acid vaccine comprising (a) at least one RNA polynucleotide, said polynucleotide comprising at least one chemical modification or optionally no modified nucleotides and two or more codon-optimized open reading frames, said open reading frames encoding a set of reference antigenic polypeptides, and (b) optionally a pharmaceutically acceptable carrier or excipient. In some embodiments, the vaccine is administered to the individual via a route selected from the group consisting of intramuscular administration, intradermal administration and subcutaneous administration. In some embodiments, the administering step comprises contacting a muscle tissue of the subject with a device suitable for injection of the composition. In some embodiments, the administering step comprises contacting a muscle tissue of the subject with a device suitable for injection of the composition in combination with electroporation.

Aspects of the invention provide methods of vaccinating a subject comprising administering to the subject a single dosage of between 25 ug/kg and 400 ug/kg of a nucleic acid vaccine comprising one or more RNA polynucleotides having an open reading frame encoding a first antigenic polypeptide in an effective amount to vaccinate the subject.

Other aspects provide nucleic acid vaccines comprising one or more RNA polynucleotides having an open reading frame comprising at least one chemical modification, the open reading frame encoding a first antigenic polypeptide, wherein the vaccine has at least 10 fold less RNA polynucleotide than is required for an unmodified mRNA vaccine to produce an equivalent antibody titer. In some embodiments, the RNA polynucleotide is present in a dosage of 25-100 micrograms.

Other aspects provide nucleic acid vaccines comprising an LNP formulated RNA polynucleotide having an open reading frame comprising no nucleotide modifications (unmodified), the open reading frame encoding a first antigenic polypeptide, wherein the vaccine has at least 10 fold less RNA polynucleotide than is required for an unmodified mRNA vaccine not formulated in a LNP to produce an equivalent antibody titer. In some embodiments, the RNA polynucleotide is present in a dosage of 25-100 micrograms.

The data presented in the Examples demonstrate significant enhanced immune responses using the formulations of the invention. Both chemically modified and unmodified RNA vaccines are useful according to the invention. Surprisingly, in contrast to prior art reports that it was preferable to use chemically unmodified mRNA formulated in a carrier for the production of vaccines, it is described herein that chemically modified mRNA-LNP vaccines required a much lower effective mRNA dose than unmodified mRNA, i.e., tenfold less than unmodified mRNA when formulated in carriers other than LNP. Both the chemically modified and unmodified RNA vaccines of the invention produce better immune responses than mRNA vaccines formulated in a different lipid carrier.

In other aspects the invention encompasses a method of treating an elderly subject age 60 years or older comprising administering to the subject a nucleic acid vaccine comprising one or more RNA polynucleotides having an open reading frame encoding a virus antigenic polypeptide in an effective amount to vaccinate the subject.

In other aspects the invention encompasses a method of treating a young subject age 17 years or younger comprising administering to the subject a nucleic acid vaccine comprising one or more RNA polynucleotides having an open reading frame encoding a virus antigenic polypeptide in an effective amount to vaccinate the subject.

In other aspects the invention encompasses a method of treating an adult subject comprising administering to the subject a nucleic acid vaccine comprising one or more RNA polynucleotides having an open reading frame encoding a virus antigenic polypeptide in an effective amount to vaccinate the subject.

In some aspects the invention is a method of vaccinating a subject with a combination vaccine including at least two nucleic acid sequences encoding antigens wherein the dosage for the vaccine is a combined therapeutic dosage wherein the dosage of each individual nucleic acid encoding an antigen is a sub therapeutic dosage. In some embodiments, the combined dosage is 25 micrograms of the RNA polynucleotide in the nucleic acid vaccine administered to the subject. In some embodiments, the combined dosage is 100 micrograms of the RNA polynucleotide in the nucleic acid vaccine administered to the subject. In some embodiments the combined dosage is 50 micrograms of the RNA polynucleotide in the nucleic acid vaccine administered to the subject. In some embodiments, the combined dosage is 75 micrograms of the RNA polynucleotide in the nucleic acid vaccine administered to the subject. In some embodiments, the combined dosage is 150 micrograms of the RNA polynucleotide in the nucleic acid vaccine administered to the subject. In some embodiments, the combined dosage is 400 micrograms of the RNA polynucleotide in the nucleic acid vaccine administered to the subject. In some embodiments, the sub therapeutic dosage of each individual nucleic acid encoding an antigen is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 micrograms. In other embodiments the nucleic acid vaccine is chemically modified and in other embodiments the nucleic acid vaccine is not chemically modified.

The RNA polynucleotide is one of SEQ ID NO: 1-12, 18-21, 30-39, 48-51, 55-58, 56, 65-96, 118-155, 223-256 or 376-401 and includes at least one chemical modification. In other embodiments the RNA polynucleotide is one of SEQ ID NO: 1-12, 18-21, 30-39, 48-51, 55-58, 56, 65-96, 118-155, 223-256 or 376-401 and does not include any nucleotide modifications, or is unmodified. In yet other embodiments the at least one RNA polynucleotide encodes an antigenic protein of any of SEQ ID NO: 13-17, 22-29, 44-47, 52-54, 59-64, 97-117, 156-222, 469, 259-291 or 402-413 and includes at least one chemical modification. In other embodiments the RNA polynucleotide encodes an antigenic protein of any of SEQ ID NO: 13-17, 22-29, 44-47, 52-54, 59-64, 97-117, 156-222, 469, 259-291 or 402-413 and does not include any nucleotide modifications, or is unmodified.

In preferred aspects, vaccines of the invention (e.g., LNP-encapsulated mRNA vaccines) produce prophylactically- and/or therapeutically-efficacious levels, concentrations and/or titers of antigen-specific antibodies in the blood or serum of a vaccinated subject. As defined herein, the term antibody titer refers to the amount of antigen-specific antibody produces in s subject, e.g., a human subject. In exemplary embodiments, antibody titer is expressed as the inverse of the greatest dilution (in a serial dilution) that still gives a positive result. In exemplary embodiments, antibody titer is determined or measured by enzyme-linked immunosorbent assay (ELISA). In exemplary embodiments, antibody titer is determined or measured by neutralization assay, e.g., by microneutralization assay. In certain aspects, antibody titer measurement is expressed as a ratio, such as 1:40, 1:100, etc.

In exemplary embodiments of the invention, an efficacious vaccine produces an antibody titer of greater than 1:40, greater that 1:100, greater than 1:400, greater than 1:1000, greater than 1:2000, greater than 1:3000, greater than 1:4000, greater than 1:500, greater than 1:6000, greater than 1:7500, greater than 1:10000. In exemplary embodiments, the antibody titer is produced or reached by 10 days following vaccination, by 20 days following vaccination, by 30 days following vaccination, by 40 days following vaccination, or by 50 or more days following vaccination. In exemplary embodiments, the titer is produced or reached following a single dose of vaccine administered to the subject. In other embodiments, the titer is produced or reached following multiple doses, e.g., following a first and a second dose (e.g., a booster dose.)

In exemplary aspects of the invention, antigen-specific antibodies are measured in units of μg/ml or are measured in units of IU/L (International Units per liter) or mIU/ml (milli International Units per ml). In exemplary embodiments of the invention, an efficacious vaccine produces >0.5 μg/ml, >0.1 μg/ml, >0.2 μg/ml, >0.35 μg/ml, >0.5 μg/ml, >1 μg/ml, >2 μg/ml, >5 μg/ml or >10 μg/ml. In exemplary embodiments of the invention, an efficacious vaccine produces >10 mIU/ml, >20 mIU/ml, >50 mIU/ml, >100 mIU/ml, >200 mIU/ml, >500 mIU/ml or >1000 mIU/ml. In exemplary embodiments, the antibody level or concentration is produced or reached by 10 days following vaccination, by 20 days following vaccination, by 30 days following vaccination, by 40 days following vaccination, or by 50 or more days following vaccination. In exemplary embodiments, the level or concentration is produced or reached following a single dose of vaccine administered to the subject. In other embodiments, the level or concentration is produced or reached following multiple doses, e.g., following a first and a second dose (e.g., a booster dose.) In exemplary embodiments, antibody level or concentration is determined or measured by enzyme-linked immunosorbent assay (ELISA). In exemplary embodiments, antibody level or concentration is determined or measured by neutralization assay, e.g., by microneutralization assay.

The details of various embodiments of the disclosure are set forth in the description below. Other features, objects, and advantages of the disclosure will be apparent from the description and from the claims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows data from an immunogenicity experiment in which mice were immunized with JEV prME mRNA vaccine. The data show that immunization of mice with JEV mRNA vaccine at 10 μg, 2 μg and 0.5 μg doses produces neutralizing antibodies measured between 10² to 10⁴ PRNT50 titers.

FIG. 2 shows a histogram indicating intracellular detection of ZIKA prME protein using human serum containing anti-ZIKV antigen antibodies.

FIG. 3 shows the results of detecting prME protein expression in mammalian cells with fluorescence-activated cell sorting (FACS) using a flow cytometer. Cells expressing prME showed higher fluorescence intensity when stained with anti-ZIKV human serum.

FIG. 4 shows a bar graph of the data provided in FIG. 3.

FIG. 5 shows a reducing SDS-PAGE gel of Zika VLP.

FIG. 6 shows a graph of neutralizing titers obtained from BALB/c mice immunized with a ZIKV mRNA vaccine encoding prME.

FIGS. 7A-7B show percent animal survival (FIG. 7A) and percent weight change (FIG. 7B) in animals following administration of two different doses of a ZIKV RNA vaccine comprising mRNA encoding ZIKV prME.

FIGS. 8A-8C show Dengue Virus MHC I T cell epitopes. The sequences, from left to right correspond to SEQ ID NO: 365-366 (FIG. 8A), 367-368 (FIG. 8B), and 369-370 (FIG. 8C).

FIGS. 9A-9C show Dengue Virus MHC II T cell epitopes. The sequences, from left to right, correspond to SEQ ID NO: 371-372 (FIG. 9A), 373-374 (FIG. 9B), and 375 (FIG. 9C).

FIG. 10 is a graph depicting the results of an ELISPOT assay of dengue-specific peptides.

FIG. 11 is a graph depicting the results of an ELISPOT assay of dengue-specific peptides.

FIG. 12 is a schematic of a bone marrow/liver/thymus (BLT) mouse and data on human CD8 T cells stimulated with Dengue peptide epitope.

FIGS. 13A and 13B shows the results of an Intracellular Cytokine Staining assay performed in PBMC cells.

FIG. 14A shows FACS analyses of cells expressing DENV2 prMEs using different antibodies against Dengue envelope protein. Numbers in the upper right corner of each plot indicate mean fluorescent intensity. FIG. 14B shows a repeat of staining in triplicate and in two different cell lines (HeLa and 293T).

FIG. 15 is a graph showing the kinetics of OVA peptide presentation in Jawsii cells. All mRNAs tested are formulated in MC3 lipid nanoparticles.

FIG. 16 is a graph showing the Mean Fluorescent Intensity (MFI) of antibody binding to DENV-1, 2, 3, and 4 prME epitopes presented on the cell surface.

FIGS. 17A-17D are graphs showing the design and the results of a challenge study in AG129 mice. FIG. 17A shows the immunization, challenge, and serum collection schedules. FIG. 17B shows the survival of the AG129 mice challenged with Dengue D2Y98P virus after being immunized with the indicated DENV mRNA vaccines. All immunized mice survived 11 days post infection, while the unimmunized (control) mice died. FIGS. 17C and 17D show the weight loss of the AG129 mice post infection. Vaccine 1, 7, 8, or 9 correspond to DENV vaccine construct 22, 21, 23, or 24 of the present disclosure, respectively.

FIG. 18 is a graph showing the results of an in vitro neutralization assay using serum from mice immunized with the DENV mRNA vaccines in FIGS. 17A-17D.

FIGS. 19A-19I are graphs showing the results of a challenge study in AG129 mice. The challenge study design is shown in Table 46. FIGS. 19A-19F show the survival, weight loss, and heath score of the AG129 mice challenged with D2Y98P virus after being immunized with the DENV mRNA vaccine groups 1-12 in Table 46. FIGS. 19G-19I show the survival, weight loss, and heath score of the AG129 mice challenged with D2Y98P virus after being immunized with the DENV mRNA vaccine groups 13-19 in Table 46.

FIG. 20 shows CHIKV envelope protein detection of lysate in HeLa cells 16 hours post-transfection.

FIG. 21A is a graph showing the survival rates of AG129 mice vaccinated with a single 2 μg dose or two 2 μg doses of Chikungunya E1 antigen administered either intramuscularly or intradermally. FIG. 21B is a graph showing the percent weight loss of AG129 mice vaccinated with a single 2 μg dose or two 2 μg doses of Chikungunya E1 antigen administered either intramuscularly or intradermally. FIG. 21C is a graph showing the health scores of AG129 mice vaccinated with a single 2 μg dose or two 2 μg doses of Chikungunya E1 antigen administered either intramuscularly or intradermally.

FIG. 22A is a graph showing the survival rates of AG129 mice vaccinated with a single 2 μg dose or two 2 μg doses of Chikungunya E2 antigen administered either intramuscularly or intradermally. FIG. 22B is a graph showing the percent weight loss of AG129 mice vaccinated with a single 2 μg dose or two 2 μg doses of Chikungunya E2 antigen administered either intramuscularly or intradermally. FIG. 22C is a graph showing the health scores of AG129 mice vaccinated with a single 2 μg dose or two 2 μg doses of Chikungunya E2 antigen administered either intramuscularly or intradermally.

FIG. 23A is a graph showing the survival rates of AG129 mice vaccinated with a single 2 μg dose or two 2 μg doses of Chikungunya C-E3-E2-6K-E1 antigen administered either intramuscularly or intradermally. FIG. 23B is a graph showing the percent weight loss of AG129 mice vaccinated with a single 2 μg dose or two 2 μg doses of Chikungunya C-E3-E2-6K-E1 antigen administered either intramuscularly or intradermally. FIG. 23C is a graph showing the health scores of AG129 mice vaccinated with a single 2 μg dose or two 2 μg doses of Chikungunya C-E3-E2-6K-E1 antigen administered either intramuscularly or intradermally.

FIG. 24A is a graph showing the survival rates of AG129 mice vaccinated with a single 10 μg dose or two 10 μg doses of Chikungunya E1 antigen administered either intramuscularly or intradermally. FIG. 24B is a graph showing the percent weight loss of AG129 mice vaccinated with a single 10 μg dose or two 10 μg doses of Chikungunya E1 antigen administered either intramuscularly or intradermally. FIG. 24C is a graph showing the health scores of AG129 mice vaccinated with a single 10 μg dose or two 10 μg doses of Chikungunya E1 antigen administered either intramuscularly or intradermally.

FIG. 25A is a graph showing the survival rates of AG129 mice vaccinated with a single 10 μg dose or two 10 μg doses of Chikungunya E2 antigen administered either intramuscularly or intradermally. FIG. 25B is a graph showing the percent weight loss of AG129 mice vaccinated with a single 10 μg dose or two 10 μg doses of Chikungunya E2 antigen administered either intramuscularly or intradermally. FIG. 25C is a graph showing the health scores of AG129 mice vaccinated with a single 10 μg dose or two 10 μg doses of Chikungunya E2 antigen administered either intramuscularly or intradermally.

FIG. 26A is a graph showing the survival rates of AG129 mice vaccinated with a single 10 μg dose or two 10 μg doses of Chikungunya C-E3-E2-6K-E1 antigen administered either intramuscularly or intradermally. FIG. 26B is a graph showing the percent weight loss of AG129 mice vaccinated with a single 10 μg dose or two 10 μg doses of Chikungunya C-E3-E2-6K-E1 antigen administered either intramuscularly or intradermally. FIG. 26C is a graph showing the health scores of AG129 mice vaccinated with a single 10 μg dose or two 10 μg doses of Chikungunya C-E3-E2-6K-E1 antigen administered either intramuscularly or intradermally.

FIGS. 27A-27B are graphs showing the survival curves from a CHIKV challenge study in AG129 mice immunized with CHIKV mRNA vaccines in 10 μg, 2 μg, or 0.04 μg doses. Mice were divided into 14 groups (1-4 and 7-16, n=5). FIG. 27A shows the survival curve of mice groups 1-4 and 7-9 challenged on day 56 post immunization. FIG. 27B shows the survival curve of mice groups 10-16 challenged on day 112 post immunization. Survival curves were plotted as “percent survival” versus “days post infection.” See also Table 63 for survival percentage.

FIGS. 28A-28B are graphs showing the weight changes post challenge in AG129 mice immunized with CHIKV mRNA vaccines. FIG. 28A shows the weight change of mice groups 1-4 and 7-9 challenged on day 56 post immunization. FIG. 28B shows the weight changes of mice groups 10-16 challenged on day 112 post immunization. Initial weights were assessed on individual mice on study Day 0 and daily thereafter. The mean percent weights for each group compared to their percent weight on Day 0 (baseline) were plotted against “days post-infection”. Error bars represent the standard deviation (SD).

FIGS. 29A-29B are graphs showing the post challenge heath scores of AG129 mice immunized with CHIKV mRNA vaccines. FIG. 29A shows the health scores of mice groups 1-4 and 7-9 challenged on day 56 post immunization. FIG. 29B shows the health score of mice groups 10-16 challenged on day 112 post immunization. The mean health scores for each group were plotted against “days post infection” and error bars represent the SD. Mean health scores were calculated based on observations described in Table 51.

FIGS. 30A-30C are graphs showing the antibody titers measured by ELISA assays in the serum of AG129 mice (groups 1-4 and 7-9) 28 days post immunization with CHIKV mRNA vaccines. FIG. 30A shows the serum antibody titers against CHIKV E1 protein. FIG. 30B shows the serum antibody titers against CHIKV E2 protein. FIG. 30C shows the serum antibody titers against CHIKV lysate.

FIGS. 31A-31C are graphs showing the antibody titers measured by ELISA assays in the serum of AG129 mice (groups 10-16) 28 days post immunization with CHIKV mRNA vaccine. FIG. 31A shows the serum antibody titers against CHIKV E1 protein. FIG. 31B shows the serum antibody titers against CHIKV E2 protein. FIG. 31C shows the serum antibody titers against CHIKV lysate.

FIGS. 32A-32C are graphs showing the antibody titers measured by ELISA assays in the serum of AG129 mice (groups 10-16) 56 days post immunization with CHIKV mRNA vaccine. FIG. 32A shows the serum antibody titers against CHIKV E1 protein. FIG. 32B shows the serum antibody titers against CHIKV E2 protein. FIG. 32C shows the serum antibody titers against CHIKV lysate.

FIGS. 33A-33C are graphs showing the antibody titers measured by ELISA assays in the serum of AG129 mice (groups 10-16) 112 days post immunization with CHIKV mRNA vaccine. FIG. 33A shows the serum antibody titers against CHIKV E1 protein. FIG. 33B shows the serum antibody titers against CHIKV E2 protein. FIG. 33C shows the serum antibody titers against CHIKV lysate.

FIG. 34 shows a set of graphs depicting results of an ELISA assay to identify the amount of antibodies produced in AG129 mice in response to vaccination with mRNA encoding secreted CHIKV E1 structural protein, secreted CHIKV E2 structural protein, or CHIKV full structural polyprotein C-E3-E2-6k-E1 at a dose of 10 μg or 2 μg at 28 days post immunization.

FIG. 35 shows a set of graphs depicting results of an ELISA assay to identify the amount of antibodies produced in AG129 mice in response to vaccination with mRNA encoding secreted CHIKV E1 structural protein, secreted CHIKV E2 structural protein, or CHIKV full structural polyprotein C-E3-E2-6k-E1 at a dose of 10 μg or 2 μg at 28 days post immunization. The two panels depict different studies.

FIG. 36 is a graph depicting comparison of ELISA titers from the data of FIG. 34 to survival in the data of FIG. 35 left panel.

FIG. 37 shows a set of graphs depicting efficacy results in mice in response to vaccination with mRNA encoding CHIKV full structural polyprotein C-E3-E2-6k-E1 at a dose of 10 μg (left panels), 2 μg (middle panels) or 0.4 μg (right panels) at 56 days (top panels) or 112 days (bottom panels) post-immunization.

FIG. 38 shows a set of graphs depicting amount of neutralizing antibody produced in mice in response to vaccination with mRNA encoding CHIKV full structural polyprotein C-E3-E2-6k-E1 at a dose of 10 μg, 2 μg, or 0.4 μg at 56 days post immunization.

FIG. 39 shows a set of graphs depicting binding antibody produced in mice in response to vaccination with mRNA encoding CHIKV full structural polyprotein C-E3-E2-6k-E1 at a dose of 10 μg, 2 μg, or 0.4 μg at 56 days post immunization (top panels) and the corresponding correlation between binding and neutralizing antibodies (bottom panels).

FIG. 40 shows a set of graphs depicting amount of neutralizing antibody produced in A129 mice in response to vaccination with mRNA encoding CHIKV full structural polyprotein C-E3-E2-6k-E1 at a dose of 10 μg, 2 μg, or 0.4 μg at 56 days post immunization against three different strains of CHIKV, African—Senegal (left panel), La Reunion (middle panel) and CDC CAR (right panel).

FIG. 41 shows a graph depicting neutralizing antibodies against CHIKV S27 strain.

FIG. 42 is a graph depicting antibody titer against CHIKV lysate post 3rd vaccination 10 with the mRNA vaccine in Sprague Dawley rats.

FIG. 43 shows a set of graphs depicting antibody titers following vaccination of mice with mRNA encoded CHIKV polyprotein (C-E3-E2-6K-E1).

FIG. 44 shows a set of plots depicting cytokine secretion and T-cell activation following vaccination of mice with mRNA encoded CHIKV polyprotein (C-E3-E2-6K-E1).

FIGS. 45A-45B show a set of graphs depicting CD8+ T cell activation following vaccination of mice with mRNA encoded CHIKV polyprotein (C-E3-E2-6K-E1).

FIG. 46 shows a set of graphs depicting binding antibody titers against CHIKV lysate (upper graph) and neutralizing titers against 37997 CHIKV. The vaccine induces a robust antibody response in non-human primates (NHPs).

FIG. 47 shows a set of graphs depicting a robust CD4 response to a CHIKV vaccine in NHPs.

DETAILED DESCRIPTION

Vaccines containing antigens from more than one pathogenic organism within a single dose are referred to as “multivalent” or “combination” vaccines. While various combination vaccines have been approved for human use in several countries, including trivalent vaccines for protecting against diphtheria, tetanus and pertussis (“DTP” vaccines) and trivalent vaccines for protecting against measles, mumps and rubella (“MMR” vaccines), combination vaccines are more complex and are associated with more problems than monovalent vaccines. For instance, current combination vaccines can include relatively high amounts of aluminum salts as adjuvants which causes concern to some patients despite empirical safety studies. Additionally, the well-documented phenomenon of antigenic competition (or interference) complicates the development of multi-component vaccines. Antigenic interference refers to the observation that administering multiple antigens often results in a diminished response to certain antigens relative to the immune response observed when such antigens are administered individually. The combination RNA vaccines of the invention can be designed to encode two, three, four, five or more, antigens against multiple pathogenic organisms, while avoiding a number of the problems associated with traditional combination vaccines.

Travelers facing a particular geographic viral threat would also benefit from vaccination with a combination vaccine of the invention. The traveler's vaccine may be tailored based on the prevalence of particular viral diseases in the destination location. For instance a combination vaccine including WNV, SINV, VEEV, and EEEV would be particularly beneficial.

Embodiments of the present disclosure provide RNA (e.g., mRNA) vaccines that are useful for vaccinating against multiple pathogens. The combination vaccines of the present disclosure encode antigens from multiple pathogens (e.g., bacteria, arboviruses, alphaviruses and flaviviruses), including but not limited to Plasmodium (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), Japanese Encephalitis Virus (JEV), West Nile Virus (WNV), Eastern Equine Encephalitis (EEEV), Venezuelan Equine Encephalitis Virus (VEEV), Sindbis Virus (SINV), Chikungunya Virus (CHIKV), Dengue Virus (DENV), Zika Virus (ZIKV) and/or Yellow Fever Virus (YFV) antigenic polypeptide.

Thus, the present disclosure provides, in some embodiments, vaccines that comprise RNA (e.g., mRNA) polynucleotides encoding a Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide. The present disclosure also provides, in some embodiments, combination vaccines that comprise at least one RNA (e.g., mRNA) polynucleotide encoding at least two antigenic polypeptides selected from Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and YFV antigenic polypeptides. Also provided herein are methods of administering the RNA (e.g., mRNA) vaccines, methods of producing the RNA (e.g., mRNA) vaccines, compositions (e.g., pharmaceutical compositions) comprising the RNA (e.g., mRNA) vaccines, and nucleic acids (e.g., DNA) encoding the RNA (e.g., mRNA) vaccines. In some embodiments, a RNA (e.g., mRNA) vaccine comprises an adjuvant, such as a flagellin adjuvant, as provided herein.

The RNA (e.g., mRNA) vaccines (e.g., Malaria (e.g., P. falciparum, P. vivax, P. Malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV RNA vaccines), in some embodiments, may be used to induce a balanced immune response, comprising both cellular and humoral immunity, without many of the risks associated with DNA vaccination.

The entire contents of International Application No. PCT/US2015/02740 is incorporated herein by reference.

Malaria

Malaria is an infectious disease caused by protozoan parasites from the Plasmodium family. Anopheles mosquitoes transmit Malaria, and they must have been infected through a previous blood meal taken from an infected person. When a mosquito bites an infected person, a small amount of blood is taken in and contains microscopic Malaria parasites. There are four main types of Malaria which infect humans: Plasmodium falciparum, P. vivax, P. Malariae and P. ovale. Falciparum Malaria is the most deadly type. Many Malaria parasites are now immune to the most common drugs used to treat the disease.

Embodiments of the present disclosure provide RNA (e.g., mRNA) vaccines that include polynucleotide encoding a Plasmodium antigen. Malaria parasites are microorganisms that belong to the genus Plasmodium. There are more than 100 species of Plasmodium, which can infect many animal species such as reptiles, birds, and various mammals. Four species of Plasmodium have long been recognized to infect humans in nature, including P. falciparum, P. vivax, P. Malariae and P. ovale. In addition, there is one species that naturally infects macaques which has recently been recognized to be a cause of zoonotic Malaria in humans.

Malaria RNA (e.g., mRNA) vaccines, as provided herein may be used to induce a balanced immune response, comprising both cellular and humoral immunity, without many of the risks associated with DNA vaccination.

P. falciparum infects humans and is found worldwide in tropical and subtropical areas. It is estimated that every year approximately 1 million people are killed by P. falciparum, especially in Africa where this species predominates. P. falciparum can cause severe Malaria because it multiples rapidly in the blood, and can thus cause severe blood loss (anemia). In addition, the infected parasites can clog small blood vessels. When this occurs in the brain, cerebral Malaria results, a complication that can be fatal. Some embodiments of the present disclosure provide Malaria vaccines that include at least one RNA (e.g., mRNA) polynucleotide having an open reading frame encoding at least one P. falciparum antigenic polypeptide or an immunogenic fragment thereof (e.g., an immunogenic fragment capable of raising an immune response to P. falciparum).

P. vivax infects humans and is found mostly in Asia, Latin America, and in some parts of Africa. Because of the population densities, especially in Asia, it is probably the most prevalent human Malaria parasite. P. vivax (as well as P. ovale) has dormant liver stages (“hypnozoites”) that can activate and invade the blood (“relapse”) several months or years after the infecting mosquito bite. Some embodiments of the present disclosure provide Malaria vaccines that include at least RNA (e.g., mRNA) polynucleotide having an open reading frame encoding at least one P. vivax antigenic polypeptide or an immunogenic fragment thereof (e.g., an immunogenic fragment capable of raising an immune response to P. vivax).

P. ovale infects humans and is found mostly in Africa (especially West Africa) and the islands of the western Pacific. It is biologically and morphologically very similar to P. vivax. However, differently from P. vivax, it can infect individuals who are negative for the Duffy blood group, which is the case for many residents of sub-Saharan Africa. This explains the greater prevalence of P. ovale (rather than P. vivax) in most of Africa. Some embodiments of the present disclosure provide Malaria vaccines that include at least one RNA (e.g., mRNA) polynucleotide having an open reading frame encoding at least one P. ovale antigenic polypeptide or an immunogenic fragment thereof (e.g., an immunogenic fragment capable of raising an immune response to P. ovale).

P. Malariae infects humans and is found worldwide. It is the only human Malaria parasite species that has a quartan cycle (three-day cycle). The three other species that infect human have a tertian, two-day cycle. If untreated, P. Malariae causes a long-lasting, chronic infection that in some cases can last a lifetime. In some chronically infected patients P. Malariae can cause serious complications such as the nephrotic syndrome. Some embodiments of the present disclosure provide Malaria vaccines that include at least one RNA (e.g., mRNA) polynucleotide having an open reading frame encoding at least one P. Malariae antigenic polypeptide or an immunogenic fragment thereof (e.g., an immunogenic fragment capable of raising an immune response to P. Malariae).

P. knowlesi is found throughout Southeast Asia as a natural pathogen of long-tailed and pig-tailed macaques. It has recently been shown to be a significant cause of zoonotic Malaria in that region, particularly in Malaysia. P. knowlesi has a 24-hour replication cycle and so can rapidly progress from an uncomplicated to a severe infection; fatal cases have been reported.

In some embodiments, an antigenic polypeptide is any antigen that is expressed on the sporozoite or other pre-erythrocytic stage of a Plasmodium parasite, such as the liver stage. For example, an antigenic; polypeptide may be a circumsporozoite (CS) protein, liver stage antigen-1 (LSA1) (see, e.g., WO2004/044167 and Cummings J F et al. Vaccine 2010; 28:5135-44, incorporated herein by reference), liver stage antigen-3 (LSA-3) (see, e.g., EP 0 570 489 and EP 0 833 917, incorporated herein by reference), Pfs 16 kD (see, e.g., WO 91/18922 and EP 597 843), Exported antigen-1 (Exp-1) (described for example in Meraldi et al. Parasite Immunol 2002; 24(3):141, incorporated herein by reference), sporozoite-threonine-asparagine-rich protein (STARP), sporozoite and liver stage antigen (SALSA), thrombospondin related anonymous protein (TRAP) (see, e.g., WO 90/01496, WO 91/11516 and WO 92/11868, incorporated herein by reference) and apical merozoite antigen-1 (AMA1) (see, e.g., EP 0 372 019 and Reniargue E J et al. Trends in Parasitology 2007; 24(2):74-84, incorporated herein by reference) which has recently been shown to be present at the liver stage (in addition to the erythrocytic stage), and merozoite surface protein-1 (MSP1) (see, e.g., Reed Z H et al. Vaccine 2009; 27:1651-60, incorporated herein by reference). An antigenic polypeptide may be the entire protein, an immunogenic fragment thereof, or a derivative thereof of any of the foregoing antigens. Immunogenic fragments of Malaria antigens are known, including, for example, the ectodomain from AMA1 (see, e.g., WO 02/077195, incorporated herein by reference). Derivatives include, for example, fusions with other proteins that may be Malaria proteins or non-Malaria proteins, such as HBsAg. Derivatives of the present disclosure are capable of raising an immune response against the native antigen.

The sporozoite stage of Plasmodium (e.g., P. falciparum or P. vivax) is a potential target of a Malaria vaccine. The major surface protein of the sporozoite is circumsporozoite protein (CS protein). The Plasmodium, circumsporozoite protein (CS) is expressed during the sporozoite and early liver stages of parasitic infection. This protein is involved in the adhesion of the sporozoite to the hepatocyte and invasion of the hepatocyte. Anti-CS antibodies inhibit parasite invasion and are also associated with a reduced risk of clinical Malaria in some studies. Antibodies raised through immunization with only the conserved Asparagine-Alanine-Asparagine-Proline (NANP) amino acid repeat sequence, the immunodominant B-cell epitope from P. falciparum CS, are capable of blocking sporozoite invasion of hepatocytes.

CS protein has been cloned, expressed and sequenced for a variety of strains, for example for P. falciparum the NF54 strain, clone 3D7 (Gaspers et al. Parasitol 1989; 35:185-190, incorporated herein by reference). The protein from strain 3D7 has a central immunodominant repeat region comprising a tetrapeptide Asn-Ala-Asn-Pro (SEQ ID NO: 434) repeated 40 times and interspersed with four minor repeats of the tetrapeptide Asn-Val-Asp-Pro (SEQ ID NO: 435). In other strains, the number of major and minor repeats as well as their relative position varies. This central portion is flanked by an N and C terminal portion composed of non-repetitive amino acid sequences designated as the repeatless portion of the CS protein.

Some embodiments of the present disclosure provide Malaria vaccines that include at least one RNA (e.g., mRNA) polynucleotide having an open reading frame encoding Plasmodium CS protein or an immunogenic fragment thereof (e.g., an immunogenic fragment capable of raising an immune response to Plasmodium).

Liver Stage Antigen-1 (LSA1), expressed during Plasmodium falciparum hepatic schizogony is highly conserved, is abundantly expressed from early through late schizogony, presumably allowing time for both circulating and memory-recalled effector cells to infiltrate the liver and exert their effector function, and it is possible that high titer antibody could act upon the cloud of flocculent liver stage antigen enveloping hepatic merozoites to impede the latter's emergence and subsequent invasion of erythrocytes. LSA1 is a 230 kDa protein, with a large central repeat region (over 80 repeats of 17 amino acids each) flanked by two highly conserved N- and C-terminal regions, known to contain B cell and CD4+ and CD8⁺ T cell epitopes.

Some embodiments of the present disclosure provide Malaria vaccines that include at least one RNA (e.g., mRNA) polynucleotide having an open reading frame encoding Plasmodium LSA1 or an immunogenic fragment thereof (e.g., an immunogenic fragment capable of raising an immune response to Plasmodium). In some embodiments, Malaria vaccines include at least one RNA (e.g., mRNA) polynucleotide having an open reading frame encoding a recombinant protein with full-length C- and N-terminal flanking domains and two of the 17 amino acid repeats from the central repeat region, referred to as “LSA-NRC.”

Present on the surface of all known Plasmodium spp., merozoite surface protein 1 (MSP1) is a polypeptide of 190-230 kDa that undergoes processing during schizont rupture to produce at least four distinct fragments (83, 28-30, 38-45 and 42 kDa). Further cleavage of the carboxy-terminal 42-kDa (MSP142) fragment yields a 19-kDa fragment (MSP119), in a process that appears to be critical for merozoite invasion. Both MSP₁₄₂ and MSP₁₁₉ regions of P. falciparum are encompassed by the present disclosure.

Thus, in some embodiments, Malaria vaccines include at least one RNA (e.g., mRNA) polynucleotide having an open reading frame encoding Plasmodium MSP1 or an immunogenic fragment thereof (e.g., an immunogenic fragment capable of raising an immune response to Plasmodium).

In some embodiments, Malaria vaccines include at least one RNA (e.g., mRNA) polynucleotide having an open reading frame encoding Plasmodium MSP1, MSP3 and AMA1.

Apical membrane antigen 1 (AMA1) is a micronemal protein of apicomplexan parasites that appears to be essential during the invasion of host cells. Immune responses to Plasmodium AMA1 can have parasite-inhibitory effects, both as measured in vitro and in animal challenge models. First identified as an invariant Plasmodium knowlesi merozoite surface antigen, AMA1 is believed to be unique to apicomplexan and derives from a single essential gene present in all Plasmodium species.

Some embodiments of the present disclosure provide Malaria vaccines that include at least one RNA (e.g., mRNA) polynucleotide having an open reading frame encoding Plasmodium AMA1 or an immunogenic fragment thereof (e.g., an immunogenic fragment capable of raising an immune response to Plasmodium).

Japanese Encephalitis Virus (JEV)

Japanese encephalitis virus (JEV), a mosquito-borne flavivirus, is a common cause of encephalitis in Asia. Japanese encephalitis (JE) occurs throughout most of Asia and parts of the western Pacific. Among an estimated 35,000-50,000 annual cases, approximately 20%-30% of patients die, and 30%-50% of survivors have neurologic or psychiatric sequelae. In endemic countries, JE is primarily a disease of children. However, travel-associated JE, although rare, can occur in a wide portion of the population. JEV is transmitted in an enzootic cycle between mosquitoes and amplifying vertebrate hosts, primarily pigs and wading birds. JEV is transmitted to humans through the bite of an infected mosquito, primarily in rural agricultural areas. In most temperate areas of Asia, JEV transmission is seasonal, and substantial epidemics can occur.

Vaccines available for use against JEV infection include live virus inactivated by such methods as formalin treatment, as well as attenuated virus (Tsai et al., in Vaccines (Plotkin, ed.) W.B. Saunders, Philadelphia, Pa., 1994, pp. 671-713). Whole virus vaccines, although effective, do have certain problems and/or disadvantages. The viruses are cultivated in mouse brain or in cell culture using mammalian cells as the host. Such culture methods are cumbersome and expensive.

Embodiments of the present disclosure provide RNA (e.g., mRNA) vaccines that include polynucleotide encoding a JEV antigen. JEV is a small-enveloped virus with a single-stranded, plus-sense RNA genome, consisting of a single open reading frame that codes for a large polyprotein which is co- and post-translationally cleaved into three structural (capsid, C; pre-membrane, prM; and envelope, E) and seven non-structural proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B and NS5). The RNA genome of JEV has a type I cap structure at its 5′-terminus but lacks a poly(A) tail at its 3′ terminus. E protein is involved in a number of important functions related to virus infection such as receptor binding and membrane fusion. E protein has been used to raise antibodies that neutralize virus activity in vitro as well as in vivo. Additionally, sub-viral particles consisting of only the prM and the E proteins were highly effective in generating protective immune response in mice against JEV. The ability of various JEV structural and non-structural proteins to produce an immune response has been examined. (Chen, H. W., et al., 1999. J. Virol. 73:10137-10145.) In view of these and other studies it has been concluded that the E protein is an important protein for inducing protective immunity against JEV.

The full-length E protein is membrane anchored. Immunogenic fragments of the E protein can be generated by removing the anchor signal. For instance, truncated Ea protein wherein a 102-amino acid hydrophobic sequence has been removed from the C-terminus of the protein to generate a 398-amino acid Es protein for immunogenic antigenic fragments. Other immunogenic fragments include a secretory form of E protein, as opposed to the anchored protein. Thus immunogenic fragments include the truncated E protein and the secretory envelope protein (Es) of JEV. JEV antigens may also include one or more non-structural proteins selected from NS1, NS2A, NS2B, NS3, NS4A, NS4B and NS5.

Since the envelope (most external portion of a JEV particle) is the first to encounter target cells, the present disclosure encompasses antigenic polypeptides associated with the envelope as immunogenic agents. In brief, surface and membrane proteins E, Es, capsid and prM—as single antigens or in combination with or without adjuvants may be used as JEV vaccine antigens.

In some embodiments, JEV vaccines comprise RNA (e.g., mRNA) encoding JEV E antigenic polypeptides or immunogenic fragments thereof.

In some embodiments, JEV vaccines comprise RNA (e.g., mRNA) encoding JEV Es antigenic polypeptides or immunogenic fragments thereof.

In some embodiments, JEV vaccines comprise RNA (e.g., mRNA) encoding JEV capsid antigenic polypeptides or immunogenic fragments thereof.

In some embodiments, JEV vaccines comprise RNA (e.g., mRNA) encoding JEV prM antigenic polypeptides or immunogenic fragments thereof.

In some embodiments, JEV vaccines comprise RNA (e.g., mRNA) encoding JEV NS1 antigenic polypeptides or immunogenic fragments thereof.

In some embodiments, JEV vaccines comprise RNA (e.g., mRNA) encoding JEV antigenic polypeptides having at least 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98% or 99% identity with JEV E protein and has receptor binding and/or membrane fusion activity.

In some embodiments, JEV vaccines comprise RNA (e.g., mRNA) encoding JEV antigenic polypeptides having at least 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98% or 99% identity with JEV Es protein and has receptor binding and/or membrane fusion activity.

In some embodiments, JEV vaccines comprise RNA (e.g., mRNA) encoding JEV antigenic polypeptides having at least 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98% or 99% identity with JEV capsid protein having capsid activity.

In some embodiments, JEV vaccines comprise RNA (e.g., mRNA) encoding JEV antigenic polypeptides having at least 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98% or 99% identity with JEV prM protein and has activity of an immature virion.

In some embodiments, JEV vaccines comprise RNA (e.g., mRNA) encoding JEV antigenic polypeptides having at least 80%, 85%, 90%, 92%, 95%, 96%, 97%, 98% or 99% identity with JEV NS1 protein and has viral replication and pathogenicity activity.

JEV RNA vaccines, as provided herein may be used to induce a balanced immune response, comprising both cellular and humoral immunity, without many of the risks associated with DNA vaccination.

West Nile Virus (WNV), Eastern Equine Encephalitis (EEEV), Venezuelan Equine Encephalitis Virus (VEEV), and Sindbis Virus (SINV)

WNV was first isolated in the West Nile region of Uganda, Africa, and belongs to the Flaviviridae family falvivirus genus. The structure of virus particles consists of a spherical structure wherein a capsid protein (C protein) is bonded to one (+) chain RNA virus gene, and a lipid bilayer membrane surrounding the spherical structure. The lipid membrane includes two kinds of proteins: envelope protein (E protein) and membrane protein (M protein). M protein is produced as a precursor prM protein and cleaved with a protease called furin to become a mature protein. West Nile virus (WNV) is an important mosquito transmitted virus which is now native to the U.S.

West Nile fever is a systemic acute fever disease caused by infection with WNV. Occasionally, the virus invades and grows in the central nervous system to cause lethal brain meningitis. WNV is widely distributed in Africa, Middle East, part of Europe, Russia, India, and Indonesia. The virus is maintained and propagated by an infection ring. The West Nile fever virus is transmitted to birds and mammals by the bites of certain mosquitoes (e.g., Culex, Aedes, Anopheles). Direct transmission may happen from WNV infected subject to healthy subject by oral transmission (prey and transmission through colostrum) and blood/organ vectored transmission. Humans, horses and domestic animals are hosts. Recently, WNV invaded and was indigenized in the US and has expanded since then. A prevalent US strain is West Nile virus NY99-flamingo382-99 strain (Lanciotti, R. S. et al., Science, 286: 2333-2337, 1999) (GenBank Accession No. AF196835).

The WNV antigens in the combination RNA vaccine may be derived from a particular WNV strain, such as NY99 or KEN-3829 or any other strain. Additional WNV strains are known in the art. West Nile virus antigens include the following proteins and polyproteins: C (capsid), E (envelope), M (membrane), prM (Pre-membrane), NS2A, NS2B, NS3 prM-E, M-E, prM-M, prM-M-E, and NS2A-NS2B-NS3.

Eastern equine encephalitis virus (EEEV), Western equine encephalitis virus (WEEV), and Venezuelan equine encephalitis virus (VEEV) are members of the Alphavirus genus of the family Togaviridae. The genus is comprised of at least 27 different arthropod-borne RNA viruses that are found throughout much of the world. The viruses normally circulate among avian or rodent hosts through the feeding activities of a variety of mosquitoes.

EEEV causes encephalitis in humans and equines in epidemic proportions. However, EEEV causes the most severe of the arboviral encephalitides in humans, with high mortality and severe neurological sequelae in survivors (Fields Virology, 4.sup.th Ed., Chapter 30 Alphaviruses, [2002] 917-962). The virus is known to be focally endemic along much of the Atlantic and Gulf Coasts of North America. It has also been found in southern Canada, the Caribbean, Central America, the eastern part of Mexico and in large sections of South America. Inland foci exist in the Great Lakes region and South Dakota in the U.S. as well as the Amazon Basin.

The current EEEV vaccine for veterinary applications in the U.S. is a formalin-inactivated whole virus preparation derived from the PE-6 strain (Bartelloni, et al. [1970] Am J. Trop Med Hyg. 19:123-126; Marie, et al. [1970] Am J Trop Med Hyg. 19:119-122). Currently there is no human vaccine. The inactivated veterinary vaccine is poorly immunogenic, requires multiple inoculations with frequent boosters and generally results in immunity of short duration.

EEEV, SINV, JEV, and CHIKV all have single-stranded, positive sense RNA genomes. A portion of the genome encodes the viral structural proteins Capsid, E3, E2, 6K, and E1, each of which are derived by proteolytic cleavage of the product of a single open reading frame. The nucleocapsid (C) protein possesses autoproteotytic activity which cleaves the C protein from the precursor protein soon after the ribosome transits the junction between the C and E3 protein coding sequence. Subsequently, the envelope glycoproteins E2 and E1 are derived by proteolytic cleavage in association with intracellular membranes and form heterodimers. E2 initially appears in the infected cell as the precursor protein PE2, which consists of E3 and E2. After extensive glycosylation and transit through the endoplasmic reticulum and the Golgi apparatus, E3 is cleaved from E2 by the furin protease. Subsequently, the E2/E1 complex is transported to the cell surface where it is incorporated into virus budding from the plasma membrane. The envelope proteins play an important role in attachment and fusion to cells.

Sindbis Virus (SINV) is also a member of the Togaviridae family, in the alphavirus subfamily and is transmitted by mosquitoes. Sindbis fever is most common in South and East Africa, Egypt, Israel, Philippines and parts of Australia. The genome encodes four non-structural proteins at the 5′ end and the capsid and two envelope proteins at the 3′ end. The non-structural proteins are involved in genome replication and the production of new genomic RNA and a shorter sub-genomic RNA strand. The viruses assemble at the host cell surfaces and acquire their envelope through budding.

Yellow Fever Virus (YFV)

Along with other viruses in the Flaviviridae family, Yellow fever virus is enveloped and icosahedral with a non-segmented, single-stranded, positive sense RNA genome. It is most closely related to the Sepik virus and is one of the two viruses in clade VIII. In 1927, Yellow fever virus was the first human virus to be isolated. It is found in tropical areas of Africa and South America. YFV is believed to have originated in Africa and spread to South America through slave trades in the 17^(th) century. Since then, there have been Yellow fever outbreaks in the Americas, Africa and Europe. It is transmitted by mosquitoes and has been isolated from a number of species in the genus Aedes (e.g., Aedes aegypti, Aedes africanus or Aedes albopictus). Mosquitos of the genus Haemagogus and Sabethes can also serve as vectors. Studies show that the extrinsic incubation period in mosquitoes is about 10 days. Vertebrate hosts of the virus include monkeys and humans.

Forty-seven African and South American countries are either endemic for, or have regions that are endemic from, Yellow fever. It is estimated that in 2013 alone, there were 84,000 to 170,000 severe cases of yellow fever and 29,000 to 60,000 deaths associated with Yellow fever.

What is important is not only the number of cases but also the clinical manifestation of the cases. After YFV incubates in the body for about 6 days, symptoms including fever, muscle pain, backache, headache, loss of appetite, and nausea or vomiting are observed. In most cases, these symptoms disappear after about 4 days. In a small percentage of patients, a more toxic phase of the disease is observed within 24 hours of recovering from the initial symptoms. In this toxic phase, patients develop high fever, jaundice, dark urine and abdominal pain with vomiting. Half of the patients that enter the toxic phase die within 10 days.

In some embodiments, YFV vaccines comprise RNA (e.g., mRNA) encoding a YFV antigenic polypeptide having at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identity with YFV polyprotein and having YFV polyprotein activity, respectively. The YFV polyprotein is cleaved into capsid, precursor membrane, envelope, and non-structural proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B, NS5).

A protein is considered to have YFV polyprotein activity if, for example, it facilitates the attachment of the viral envelope to host receptors, mediates internalization into the host cell, and aids in fusion of the virus membrane with the host's endosomal membrane.

Zika Virus (ZIKV)

Along with other viruses in the Flaviviridae family, Zika virus is enveloped and icosahedral with a non-segmented, single-stranded, positive sense RNA genome. It is most closely related to the Spondweni virus and is one of the two viruses in the Spondweni virus Glade. The virus was first isolated in 1947 from a rhesus monkey in the Zika Forest of Uganda, Africa and was isolated for the first time from humans in 1968 in Nigeria. From 1951 through 1981, evidence of human infection was reported from other African countries such as Uganda, Tanzania, Egypt, Central African Republic, Sierra Leone and Gabon, as well as in parts of Asia including India, Malaysia, the Philippines, Thailand, Vietnam and Indonesia. It is transmitted by mosquitoes and has been isolated from a number of species in the genus Aedes—Aedes aegypti, Aedes africanus, Aedes apicoargenteus, Aedes furcifer, Aedes luteocephalus and Aedes vitattus. Studies show that the extrinsic incubation period in mosquitoes is about 10 days. The vertebrate hosts of the virus include monkeys and humans.

As of early 2016, the most widespread outbreak of Zika fever, caused by the Zika virus, is ongoing primarily in the Americas. The outbreak began in April 2015 in Brazil, and subsequently spread to other countries in South America, Central America, and the Caribbean.

The Zika virus was first linked with newborn microcephaly during the Brazil Zika virus outbreak. In 2015, there were 2,782 cases of microcephaly compared with 147 in 2014 and 167 in 2013. The Brazilian Health Ministry has reported 4783 cases of suspected microcephaly as of Jan. 30, 2016, an increase of more than 1000 cases from a week earlier. Confirmation of many of the recent cases is pending, and it is difficult to estimate how many cases went unreported before the recent awareness of the risk of virus infections.

What is important is not only the number of cases but also the clinical manifestation of the cases. Brazil is seeing severe cases of microcephaly, which are more likely to be paired with greater developmental delays. Most of what is being reported out of Brazil is microcephaly with other associated abnormalities. The potential consequence of this is the fact that there are likely to be subclinical cases where the neurological sequelae will only become evident as the children grow.

Zika virus has also been associated with an increase in a rare condition known as Guillain-Barré, where the infected individual becomes essentially paralyzed. During the Zika virus outbreak in French Polynesia, of the 74 patients which had had Zika symptoms, 42 were diagnosed with Guillain-Barré syndrome. In Brazil, 121 cases of neurological manifestations and Guillain-Barré syndrome (GBS) were reported, all cases with a history of Zika-like symptoms.

The design of preferred Zika vaccine mRNA constructs of the invention encode prME proteins from the Zika virus intended to produce significant immunogenicity. The open reading frame comprises a signal peptide (to optimize expression into the endoplasmic reticulum) followed by the Zika prME polyprotein sequence. The particular prME sequence used is from a Micronesian strain (2007) that most closely represents a consensus of contemporary strain prMEs. This construct has 99% prME sequence identity to the current Brazilian isolates.

Within the Zika family, there is a high level of homology within the prME sequence (>90%) across all strains so far isolated. The high degree of homology is also preserved when comparing the original isolates from 1947 to the more contemporary strains circulating in Brazil in 2015, suggesting that there is “drift” occurring from the original isolates. Furthermore, attenuated virus preparations have provided cross-immunization to all other strains tested, including Latin American/Asian, and African. Overall, this data suggests that cross-protection of all Zika strains is possible with a vaccine based on prME. In fact, the prM/M and E proteins of ZIKV have a very high level (99%) of sequence conservation between the currently circulating Asiatic and Brazilian viral strains.

The M and E proteins are on the surface of the viral particle. Neutralizing antibodies predominantly bind to the E protein, the preM/M protein functions as a chaperone for proper folding of E protein and prevent premature fusion of E protein within acidic compartments along the cellular secretory pathway.

Described herein are examples of ZIKV vaccine designs comprising mRNA encoding the both prM/M and E proteins or E protein alone. In some embodiments, the mRNA encodes an artificial signal peptide fused to prM protein fused to E protein. In some embodiments, the mRNA encodes an artificial signal peptide fused to E protein.

ZIKV vaccine constructs can encode the prME or E proteins from different strains, for example, Brazil_isolate_ZikaSPH2015 or ACD75819_Micronesia, having a signal peptide fused to the N-termini of the antigenic protein(s). In some embodiments, ZIKV vaccines comprise mRNAs encoding antigenic polypeptides having amino acid sequences of SEQ ID NO: 156-222 or 469.

Dengue Virus (DENV)

There is no specific treatment for DENV infection, and control of DENV by vaccination has proved elusive, in part, because the pathogenesis of DHF/DSS is not completely understood. While infection with one serotype confers lifelong homotypic immunity, it confers only short term (approximately three to six months) cross protection against heterotypic serotypes. Also, there is evidence that prior infection with one type can produce an antibody response that can intensify, or enhance, the course of disease during a subsequent infection with a different serotype. The possibility that vaccine components could elicit enhancing antibody responses, as opposed to protective responses, has been a major concern in designing and testing vaccines to protect against dengue infections.

In late 2015 and early 2016, the first dengue vaccine, Dengvaxia (CYD-TDV) by Sanofi Pasteur, was registered in several countries for use in individuals 9-45 years of age living in endemic areas. Issues with the vaccine include (1) weak protection against DENV1 and DENV2 (<60% efficacy); (2) relative risk of dengue hospitalization among children <9 years old (7.5× higher than placebo); (3) immunogenicity not sustained after 1-2 years (implying the need for a 4^(th) dose booster); and (4) lowest efficacy against DENV2, which often causes more severe conditions. This latter point is a major weakness with the Dengvaxia vaccine, signaling the need of a new, more effective vaccine effective against DENV2. Other tetravalent live-attenuated vaccines are under development in phase II and phase III clinical trials, and other vaccine candidates (based on subunit, DNA and purified inactivated virus platforms) are at earlier stages of clinical development, although the ability of these vaccine candidates to provide broad serotype protection has not been demonstrated.

Embodiments of the present disclosure provide RNA (e.g., mRNA) vaccines that include at least one RNA polynucleotide encoding a Dengue virus (DENV) antigen. Dengue virus is a mosquito-borne (Aedes aegypti/Aedes albopictus) member of the family Flaviviridae (positive-sense, single-stranded RNA virus). The dengue virus genome encodes ten genes and is translated as a single polypeptide which is cut into ten proteins: the capsid, envelope, membrane, and nonstructural proteins (NS1, NS2A, NS2B, NS3, SN4A, NS4B, and NS5 proteins). The virus' main antigen is DENV envelope (E) protein, which is a component of the viral surface and is thought to facilitate the binding of the virus to cellular receptors (Heinz et al., Virology. 1983, 126:525). There are four similar but distinct serotypes of dengue virus (DENV-1, DENV-2, DENV-3, DENV-4, and DENV-5), which result annually in an estimated 50-100 million cases of dengue fever and 500,000 cases of the more severe dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS) (Gubler et al., Adv Virus Res. 1999, 53:35-70). The four serotypes show immunological cross-reactivity, but are distinguishable in plaque reduction neutralization tests and by their respective monoclonal antibodies. The dengue virus E protein includes a serotype-specific antigenic determinant and determinants necessary for virus neutralization (Mason et al., J Gen Virol. 1990, 71:2107-2114).

After inoculation, the dendritic cells become infected and travel to lymph nodes. Monocytes and macrophages are also targeted shortly thereafter. Generally, the infected individual will be protected against homotypic reinfection for life; however, the individual will only be protected against other serotypes for a few weeks or months (Sabin, Am J Trop Med Hyg. 1952, 1:30-50). In fact, DHF/DSS is generally found in children and adults infected with a dengue virus serotype differing from their respective primary infection. Thus, it is necessary to develop a vaccine that provides immunity to all four serotypes.

The DENV E (envelope) protein is found on the viral surface and plays a role in the initial attachment of the viral particle to the host cell. Several molecules which interact with the viral E protein (ICAM3-grabbing non-integrin, CD209, Rab 5, GRP 78, and the mannose receptor) are thought to be important factors mediating attachment and viral entry.

The DENV prM (membrane) protein is important in the formation and maturation of the viral particle. The membrane protein consists of seven antiparallel β-strands stabilized by three disulfide bonds. The glycoprotein shell of the mature DENV virion consists of 180 copies each of the E protein and M protein. The immature virion comprises E and prM proteins, which form 90 heterodimer spikes on the exterior of the viral particle. The immature viral particle buds into the endoplasmic reticulum and eventually travels via the secretory pathway to the Golgi apparatus. As the virion passes through the trans-Golgi Network (TGN), it is exposed to an acidic environment which causes a conformational change in the E protein which causes it to disassociate from the prM protein and form E homodimers. During the maturation phase, the pr peptide is cleaved from the M peptide by the host protease, furin. The M protein then acts as a transmembrane protein under the E-protein shell of the mature virion. The pr peptide remains associated with the E protein until the viral particle is released into the extracellular environment, acting like a cap covering the hydrophobic fusion loop of the E protein until the viral particle has exited the cell.

The DENV NS3 is a serine protease, as well as an RNA helicase and RTPase/NTPase. The protease domain consists of six β-strands arranged into two β-barrels formed by residues 1-180 of the protein. The catalytic triad (His-51, Asp-75 and Ser-135), is found between these two β-barrels, and its activity is dependent on the presence of the NS2B cofactor which wraps around the NS3 protease domain and becomes part of the active site. The remaining NS3 residues (180-618), form the three subdomains of the DENV helicase. A six-stranded parallel β-sheet surrounded by four α-helices make up subdomains I and II, and subdomain III is composed of 4 α-helices surrounded by three shorter α-helices and two antiparallel β-strands.

Chikungunya Virus (CHIKV)

Presently, CHIKV is a re-emerging human pathogen that has now established itself in Southeast Asia and has more recently spread to Europe. The Chikungunya virus (CHIKV) was introduced into Asia around 1958, and sites of endemic transmission within Southeastern Asia, including the Indian Ocean, were observed through 1996. The CHIKV epidemic moved throughout Asia, reaching Europe and Africa in the early 2000s, and was imported via travelers to North America and South America from 2005 to 2007. Sporadic outbreaks are still occurring in several countries, such as Italy, infecting naive populations. Singapore, for instance, experienced two successive waves of Chikungunya virus outbreaks in January and August 2008. Of the two strain lineages of CHIKV, the African strain remains enzootic by cycling between mosquitoes and monkeys, but the Asian strain is transmitted directly between mosquitoes and humans. This cycle of transmission may have allowed the virus to become more pathogenic as the reservoir host was eliminated.

In humans, CHIKV causes a debilitating disease characterized by fever, headache, nausea, vomiting, fatigue, rash, muscle pain and joint pain. Following the acute phase of the illness, patients develop severe chronic symptoms lasting from several weeks to months, including fatigue, incapacitating joint pain and polyarthritis.

The re-emergence of CHIKV has caused millions of cases throughout countries around the Indian Ocean and in Southeast Asia. Specifically, India, Indonesia, Maldives, Myanmar and Thailand have reported over 1.9 million cases since 2005. Globally, human CHIKV epidemics from 2004-2011 have resulted in 1.4-6.5 million reported cases, including a number of deaths. Thus, CHIKV remains a public threat that constitutes a major public health problem with severe social and economic impact.

Despite significant morbidity and some cases of mortality associated with CHIKV infection and its growing prevalence and geographic distribution, there is currently no licensed CHIKV vaccine or antiviral approved for human use. Several potential CHIKV vaccine candidates have been tested in humans and animals with varying success.

Chikungunya virus is a small (about 60-70 nm diameter), spherical, enveloped, positive-strand RNA virus having a capsid with icosahedral symmetry. The virion consists of an envelope and a nucleocapsid. The virion RNA is infectious and serves as both genome and viral messenger RNA. The genome is a linear, ssRNA(+) genome of 11,805 nucleotides which encodes two polyproteins that are processed by host and viral proteases into non-structural proteins (nsP1, nsP2, nsP3, and RdRpnsP4) necessary for RNA synthesis (replication and transcription) and structural proteins (capsid and envelope proteins C, E3, E2, 6K, and E1) which attach to host receptors and mediate endocytosis of virus into the host cell. The E1 and E2 glycoproteins form heterodimers that associate as 80 trimeric spikes on the viral surface covering the surface evenly. The envelope glycoproteins play a role in attachment to cells. The capsid protein possesses a protease activity that results in its self-cleavage from the nascent structural protein. Following its cleavage, the capsid protein binds to viral RNA and rapidly assembles into icosahedric core particles. The resulting nucleocapsid eventually associates with the cytoplasmic domain of E2 at the cell membrane, leading to budding and formation of mature virions.

E2 is an envelope glycoprotein responsible for viral attachment to target host cell, by binding to the cell receptor. E2 is synthesized as a p62 precursor which is processed at the cell membrane prior to virion budding, giving rise to an E2-E1 heterodimer. The C-terminus of E2 is involved in budding by interacting with capsid proteins.

E1 is an envelope glycoprotein with fusion activity, which is inactive as long as E1 is bound to E2 in the mature virion. Following virus attachment to target cell and endocytosis, acidification of the endosome induces dissociation of the E1/E2 heterodimer and concomitant trimerization of the E1 subunits. The E1 trimer is fusion active and promotes the release of the viral nucleocapsid in the cytoplasm after endosome and viral membrane fusion.

E3 is an accessory protein that functions as a membrane translocation/transport signal for E1 and E2.

6K is another accessory protein involved in virus glycoprotein processing, cell permeabilization, and the budding of viral particles Like E3, it functions as a membrane transport signal for E1 and E2.

The CHIKV structural proteins have been shown to be antigenic, which proteins, fragments, and epitopes thereof are encompassed within the invention. A phylogenetic tree of Chikungunya virus strains derived from complete concatenated open reading frames for the nonstructural and structural polyproteins shows key envelope glycoprotein E1 amino acid substitutions that facilitated (Indian Ocean lineage) or prevented (Asian lineage) adaptation to Aedes albopictus. There are membrane-bound and secreted forms of E1 and E2, as well as the full length polyprotein antigen (C-E3-E2-6K-E1), which retains the protein's native conformation. Additionally, the different Chikungunya genotypes, strains and isolates can also yield different antigens, which are functional in the constructs of the invention. For example, there are several different Chikungunya genotypes: Indian Ocean, East/Central/South African (ECSA), Asian, West African, and the Brazilian isolates (ECSA/Asian). There are three main Chikungunya genotype. These are ESCA (East-South-Central Africa), Asia, and West Africa. While sometimes names differ in publications, all belong to these three geographical strains.

The entire contents of International Application No. PCT/US2015/02740 is incorporated herein by reference.

Combination Vaccines

Embodiments of the present disclosure also provide combination RNA (e.g., mRNA) vaccines. A “combination RNA (e.g., mRNA) vaccine” of the present disclosure refers to a vaccine comprising at least one (e.g., at least 2, 3, 4, 5, 6, 7, 8, 9 or 10) RNA (e.g., mRNA) polynucleotide having an open reading frame encoding a combination of at least one Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, at least one JEV antigenic polypeptide, at least one WNV antigenic polypeptide, at least one EEEV antigenic polypeptide, at least one VEEV antigenic polypeptide, at least one SINV antigenic polypeptide, at least on CHIKV antigenic polypeptide, at least one DENV antigenic polypeptide, at least one ZIKV antigenic polypeptide, at least one YFV antigenic polypeptide, or any combination of two, three, four, five, six, seven, eight, nine, ten or more of the foregoing antigenic polypeptides.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a VEEV antigenic polypeptide, a SINV antigenic polypeptide, a CHIKV antigenic polypeptide, a DENV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide and a JEV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide and a WNV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide and a EEEV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide and a VEEV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide and a SINV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide and a WNV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide and a EEEV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide and a VEEV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide and a SINV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide and a EEEV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide and a VEEV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide and a SINV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide and a VEEV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide and a SINV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a VEEV antigenic polypeptide and a SINV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a VEEV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a VEEV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a VEEV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a VEEV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a SINV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a SINV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a SINV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a SINV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a CHIKV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a CHIKV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a CHIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide and a WNV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide and a EEEV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide and a VEEV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide and a SINV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide and a EEEV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide and a VEEV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide and a SINV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a EEEV antigenic polypeptide and a VEEV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a EEEV antigenic polypeptide and a SINV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a EEEV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a EEEV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a EEEV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a EEEV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a VEEV antigenic polypeptide and a SINV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a VEEV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a VEEV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a VEEV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a VEEV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, SINV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, SINV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, SINV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, SINV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a CHIKV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a CHIKV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a CHIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide and a EEEV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide and a VEEV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide and a SINV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a EEEV antigenic polypeptide and a VEEV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a EEEV antigenic polypeptide and a SINV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a EEEV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a EEEV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a EEEV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a EEEV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a VEEV antigenic polypeptide and a SINV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a VEEV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a VEEV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a VEEV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a VEEV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a SINV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a SINV antigenic polypeptide and DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a SINV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a SINV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, CHIKV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, CHIKV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, CHIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a EEEV antigenic polypeptide and a VEEV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a EEEV antigenic polypeptide and a SINV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a EEEV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a EEEV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a EEEV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a EEEV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding WNV antigenic polypeptide, a VEEV antigenic polypeptide and a SINV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding WNV antigenic polypeptide, a VEEV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding WNV antigenic polypeptide, a VEEV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding WNV antigenic polypeptide, a VEEV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding WNV antigenic polypeptide, a VEEV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a SINV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a SINV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a SINV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a SINV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a SINV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a SINV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a SINV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a SINV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a CHIKV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a CHIKV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a CHIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a DENV antigenic polypeptide and YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a ZIKV antigenic polypeptide and YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a VEEV antigenic polypeptide and a SINV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a VEEV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a VEEV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a VEEV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a VEEV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a SINV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a SINV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a SINV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a SINV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a CHIKV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a CHIKV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a CHIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a VEEV antigenic polypeptide, a SINV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a VEEV antigenic polypeptide, a SINV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a VEEV antigenic polypeptide, a SINV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a VEEV antigenic polypeptide, a SINV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a VEEV antigenic polypeptide, a CHIKV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a VEEV antigenic polypeptide, a CHIKV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a VEEV antigenic polypeptide, a CHIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a VEEV antigenic polypeptide, a DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a VEEV antigenic polypeptide, a DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a VEEV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a SINV antigenic polypeptide, a CHIKV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a SINV antigenic polypeptide, a CHIKV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a SINV antigenic polypeptide, a CHIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a SINV antigenic polypeptide, a DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a SINV antigenic polypeptide, a DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a SINV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a CHIKV antigenic polypeptide, a DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a CHIKV antigenic polypeptide, a DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a CHIKV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a DENV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a WNV antigenic polypeptide and a EEEV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a WNV antigenic polypeptide and a VEEV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a WNV antigenic polypeptide and a SINV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a WNV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a WNV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a WNV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a WNV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a EEEV antigenic polypeptide and a VEEV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a EEEV antigenic polypeptide and a SINV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a EEEV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a EEEV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a EEEV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a EEEV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide and a VEEV antigenic polypeptide and a SINV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide and a VEEV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide and a VEEV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide and a VEEV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide and a VEEV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a SINV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a SINV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a SINV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a SINV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a CHIKV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a CHIKV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a CHIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide, a EEEV antigenic polypeptide and a VEEV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide, a EEEV antigenic polypeptide and a SINV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide, a EEEV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide, a EEEV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide, a EEEV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide, a EEEV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide, a VEEV antigenic polypeptide and a SINV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide, a VEEV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide, a VEEV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide, a VEEV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide, a VEEV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide, a SINV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide, a SINV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide, a SINV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide, a SINV antigenic polypeptide and YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide, a CHIKV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide, a CHIKV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide, a CHIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide, DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide, DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide, ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, EEEV antigenic polypeptide, a VEEV antigenic polypeptide and a SINV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, EEEV antigenic polypeptide, a VEEV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, EEEV antigenic polypeptide, a VEEV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, EEEV antigenic polypeptide, a VEEV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, EEEV antigenic polypeptide, a VEEV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a SINV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a SINV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a SINV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a SINV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a EEEV antigenic polypeptide, a CHIKV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a EEEV antigenic polypeptide, a CHIKV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a EEEV antigenic polypeptide, a CHIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a EEEV antigenic polypeptide, DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a EEEV antigenic polypeptide, DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a EEEV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a VEEV antigenic polypeptide, a SINV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a VEEV antigenic polypeptide, a SINV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a VEEV antigenic polypeptide, a SINV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a VEEV antigenic polypeptide, a SINV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a VEEV antigenic polypeptide, a CHIKV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a VEEV antigenic polypeptide, a CHIKV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a VEEV antigenic polypeptide, a CHIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a VEEV antigenic polypeptide, a DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a VEEV antigenic polypeptide, a DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a VEEV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a SINV antigenic polypeptide, a CHIKV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a SINV antigenic polypeptide, a CHIKV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a SINV antigenic polypeptide, a CHIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a SINV antigenic polypeptide, a DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a SINV antigenic polypeptide, a DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a SINV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a CHIKV antigenic polypeptide, a DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a CHIKV antigenic polypeptide, a DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a CHIKV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a DENV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a EEEV antigenic polypeptide and a VEEV antigenic.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a EEEV antigenic polypeptide and a SINV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a EEEV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a EEEV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a EEEV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a EEEV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a VEEV antigenic polypeptide and a SINV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a VEEV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a VEEV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a VEEV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a VEEV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a VEEV antigenic polypeptide and a SINV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a VEEV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a VEEV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a VEEV antigenic polypeptide and a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a VEEV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a SINV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a SINV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a SINV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a SINV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a CHIKV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a CHIKV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a CHIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a ZIKV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a EEEV antigenic polypeptide, a VEEV antigenic polypeptide and a SINV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a EEEV antigenic polypeptide, a VEEV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a EEEV antigenic polypeptide, a VEEV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a EEEV antigenic polypeptide, a VEEV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a EEEV antigenic polypeptide, a VEEV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a EEEV antigenic polypeptide, a SINV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a EEEV antigenic polypeptide, a SINV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a EEEV antigenic polypeptide, a SINV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a EEEV antigenic polypeptide, a SINV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a EEEV antigenic polypeptide, a CHIKV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a EEEV antigenic polypeptide, a CHIKV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a EEEV antigenic polypeptide, a CHIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a EEEV antigenic polypeptide, a DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a EEEV antigenic polypeptide, a DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a EEEV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a VEEV antigenic polypeptide, a SINV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a VEEV antigenic polypeptide, a SINV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a VEEV antigenic polypeptide, a SINV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a VEEV antigenic polypeptide, a SINV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a VEEV antigenic polypeptide, a CHIKV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a VEEV antigenic polypeptide, a CHIKV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a VEEV antigenic polypeptide, a CHIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a VEEV antigenic polypeptide, a DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a VEEV antigenic polypeptide, a DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a VEEV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a SINV antigenic polypeptide, a CHIKV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a SINV antigenic polypeptide, a CHIKV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a SINV antigenic polypeptide, a CHIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a SINV antigenic polypeptide, a DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a SINV antigenic polypeptide, a DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a SINV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a CHIKV antigenic polypeptide, a DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a CHIKV antigenic polypeptide, a DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a CHIKV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a DENV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a VEEV antigenic polypeptide and a SINV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a VEEV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a VEEV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a VEEV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a VEEV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a SINV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a SINV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a SINV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a SINV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a CHIKV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a CHIKV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a CHIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a VEEV antigenic polypeptide, a SINV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a VEEV antigenic polypeptide, a SINV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a VEEV antigenic polypeptide, a SINV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a VEEV antigenic polypeptide, a SINV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a VEEV antigenic polypeptide, a CHIKV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a VEEV antigenic polypeptide, a CHIKV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a VEEV antigenic polypeptide, a CHIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a VEEV antigenic polypeptide, a DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a VEEV antigenic polypeptide, a DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a VEEV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a SINV antigenic polypeptide, a CHIKV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a SINV antigenic polypeptide, a CHIKV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a SINV antigenic polypeptide, a CHIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a SINV antigenic polypeptide, a DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a SINV antigenic polypeptide, a DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a SINV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a CHIKV antigenic polypeptide, a DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a CHIKV antigenic polypeptide, a DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a CHIKV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a WNV antigenic polypeptide, a DENV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a VEEV antigenic polypeptide, a SINV antigenic polypeptide and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a VEEV antigenic polypeptide, a SINV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a VEEV antigenic polypeptide, a SINV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a VEEV antigenic polypeptide, a SINV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a VEEV antigenic polypeptide, a CHIKV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a VEEV antigenic polypeptide, a CHIKV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a VEEV antigenic polypeptide, a CHIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a VEEV antigenic polypeptide, a DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a VEEV antigenic polypeptide, a DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a VEEV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a SINV antigenic polypeptide, a CHIKV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a SINV antigenic polypeptide, a CHIKV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a SINV antigenic polypeptide, a CHIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a SINV antigenic polypeptide, a DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a SINV antigenic polypeptide, a DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a SINV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a SINV antigenic polypeptide, a DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a SINV antigenic polypeptide, a DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a EEEV antigenic polypeptide, a SINV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding VEEV antigenic polypeptide, a SINV antigenic polypeptide, a CHIKV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding VEEV antigenic polypeptide, a SINV antigenic polypeptide, a CHIKV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding VEEV antigenic polypeptide, a SINV antigenic polypeptide, a CHIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding VEEV antigenic polypeptide, a SINV antigenic polypeptide, a DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding VEEV antigenic polypeptide, a SINV antigenic polypeptide, a DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding VEEV antigenic polypeptide, a SINV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding VEEV antigenic polypeptide, a CHIKV antigenic polypeptide, a DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding VEEV antigenic polypeptide, a CHIKV antigenic polypeptide, a DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding VEEV antigenic polypeptide, a CHIKV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding VEEV antigenic polypeptide, a DENV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding SINV antigenic polypeptide, a CHIKV antigenic polypeptide, a DENV antigenic polypeptide and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding SINV antigenic polypeptide, a CHIKV antigenic polypeptide, a DENV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding SINV antigenic polypeptide, a CHIKV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding SINV antigenic polypeptide, a DENV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding CHIKV antigenic polypeptide, a DENV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a VEEV antigenic polypeptide, a SINV antigenic polypeptide, a CHIKV antigenic polypeptide, a DENV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a VEEV antigenic polypeptide, a SINV antigenic polypeptide, a CHIKV antigenic polypeptide, a DENV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a EEEV antigenic polypeptide, a VEEV antigenic polypeptide, a SINV antigenic polypeptide, a CHIKV antigenic polypeptide, a DENV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a WNV antigenic polypeptide, a VEEV antigenic polypeptide, a SINV antigenic polypeptide, a CHIKV antigenic polypeptide, a DENV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a SINV antigenic polypeptide, a CHIKV antigenic polypeptide, a DENV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a VEEV antigenic polypeptide, a CHIKV antigenic polypeptide, a DENV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a VEEV antigenic polypeptide, a SINV antigenic polypeptide, a DENV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a VEEV antigenic polypeptide, a SINV antigenic polypeptide, a CHIKV antigenic polypeptide, a ZIKV antigenic polypeptide and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a VEEV antigenic polypeptide, a SINV antigenic polypeptide, a CHIKV antigenic polypeptide, a DENV antigenic polypeptide, and a YFV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide, a JEV antigenic polypeptide, a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a VEEV antigenic polypeptide, a SINV antigenic polypeptide, a CHIKV antigenic polypeptide, a DENV antigenic polypeptide, and a ZIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a VEEV antigenic polypeptide, and a SINV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a VEEV antigenic polypeptide, a SINV antigenic polypeptide, and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a SINV antigenic polypeptide, and a CHIKV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises a RNA (e.g., mRNA) polynucleotide encoding a JEV antigenic polypeptide, a WNV antigenic polypeptide, a EEEV antigenic polypeptide, a SINV antigenic polypeptide, a CHIKV antigenic polypeptide and a DENV antigenic polypeptide.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises at least two RNA (e.g., mRNA) polynucleotides selected from Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptides, JEV antigenic polypeptides, WNV antigenic polypeptides, EEEV antigenic polypeptides, VEEV antigenic polypeptides, SINV antigenic polypeptides, CHIKV antigenic polypeptides, DENV antigenic polypeptides, ZIKV antigenic polypeptides and a YFV antigenic polypeptides.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises at least three RNA (e.g., mRNA) polynucleotides selected from Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptides, JEV antigenic polypeptides, WNV antigenic polypeptides, EEEV antigenic polypeptides, VEEV antigenic polypeptides, SINV antigenic polypeptides, CHIKV antigenic polypeptides, DENV antigenic polypeptides, ZIKV antigenic polypeptides and a YFV antigenic polypeptides.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises at least four RNA (e.g., mRNA) polynucleotides selected from Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptides, JEV antigenic polypeptides, WNV antigenic polypeptides, EEEV antigenic polypeptides, VEEV antigenic polypeptides, SINV antigenic polypeptides, CHIKV antigenic polypeptides, DENV antigenic polypeptides, ZIKV antigenic polypeptides and a YFV antigenic polypeptides.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises at least five RNA (e.g., mRNA) polynucleotides selected from Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptides, JEV antigenic polypeptides, WNV antigenic polypeptides, EEEV antigenic polypeptides, VEEV antigenic polypeptides, SINV antigenic polypeptides, CHIKV antigenic polypeptides, DENV antigenic polypeptides, ZIKV antigenic polypeptides and a YFV antigenic polypeptides.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises at least six RNA (e.g., mRNA) polynucleotides selected from Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptides, JEV antigenic polypeptides, WNV antigenic polypeptides, EEEV antigenic polypeptides, VEEV antigenic polypeptides, SINV antigenic polypeptides, CHIKV antigenic polypeptides, DENV antigenic polypeptides, ZIKV antigenic polypeptides and a YFV antigenic polypeptides.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises at least seven RNA (e.g., mRNA) polynucleotides selected from Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptides, JEV antigenic polypeptides, WNV antigenic polypeptides, EEEV antigenic polypeptides, VEEV antigenic polypeptides, SINV antigenic polypeptides, CHIKV antigenic polypeptides, DENV antigenic polypeptides, ZIKV antigenic polypeptides and a YFV antigenic polypeptides.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises at least eight RNA (e.g., mRNA) polynucleotides selected from Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptides, JEV antigenic polypeptides, WNV antigenic polypeptides, EEEV antigenic polypeptides, VEEV antigenic polypeptides, SINV antigenic polypeptides, CHIKV antigenic polypeptides, DENV antigenic polypeptides, ZIKV antigenic polypeptides and a YFV antigenic polypeptides.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises at least nine RNA (e.g., mRNA) polynucleotides selected from Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptides, JEV antigenic polypeptides, WNV antigenic polypeptides, EEEV antigenic polypeptides, VEEV antigenic polypeptides, SINV antigenic polypeptides, CHIKV antigenic polypeptides, DENV antigenic polypeptides, ZIKV antigenic polypeptides and a YFV antigenic polypeptides.

In some embodiments, a combination RNA (e.g., mRNA) vaccine comprises at least ten RNA (e.g., mRNA) polynucleotides selected from Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptides, JEV antigenic polypeptides, WNV antigenic polypeptides, EEEV antigenic polypeptides, VEEV antigenic polypeptides, SINV antigenic polypeptides, CHIKV antigenic polypeptides, DENV antigenic polypeptides, ZIKV antigenic polypeptides and a YFV antigenic polypeptides.

Additional combination vaccines are encompassed by the following numbered paragraphs:

1. A combination vaccine comprising at least one RNA (e.g., mRNA) encoding at least one tropical disease antigenic polypeptide.

2. The combination vaccine of paragraph 1, wherein the at least one polypeptide is at least one Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale) antigenic polypeptide.

3. The combination vaccine of paragraph 1 or 2, wherein the at least one polypeptide is at least one JEV antigenic polypeptide.

4. The combination vaccine of any one of paragraphs 1-3, wherein the at least one polypeptide is at least one WNV antigenic polypeptide.

5. The combination vaccine of any one of paragraphs 1-4, wherein the at least one polypeptide is at least one EEEV antigenic polypeptide.

6. The combination vaccine of any one of paragraphs 1-5, wherein the at least one polypeptide is at least one VEEV antigenic polypeptide.

7. The combination vaccine of any one of paragraphs 1-6, wherein the at least one polypeptide is at least one SINV antigenic polypeptide.

8. The combination vaccine of any one of paragraphs 1-7, wherein the at least one polypeptide is at least one CHIKV antigenic polypeptide.

9. The combination vaccine of any one of paragraphs 1-8, wherein the at least one polypeptide is at least one DENV antigenic polypeptide.

10. The combination vaccine of any one of paragraphs 1-9, wherein the at least one polypeptide is at least one ZIKV antigenic polypeptide.

11. The combination vaccine of any one of paragraphs 1-10, wherein the at least one polypeptide is at least one YFV antigenic polypeptide.

It has been discovered that the mRNA vaccines described herein are superior to current vaccines in several ways. First, the lipid nanoparticle (LNP) delivery is superior to other formulations including a protamine base approach described in the literature and no additional adjuvants are to be necessary. The use of LNPs enables the effective delivery of chemically modified or unmodified mRNA vaccines. Additionally it has been demonstrated herein that both modified and unmodified LNP formulated mRNA vaccines were superior to conventional vaccines by a significant degree. In some embodiments the mRNA vaccines of the invention are superior to conventional vaccines by a factor of at least 10 fold, 20 fold, 40 fold, 50 fold, 100 fold, 500 fold or 1,000 fold.

Although attempts have been made to produce functional RNA vaccines, including mRNA vaccines and self-replicating RNA vaccines, the therapeutic efficacy of these RNA vaccines has not yet been fully established. Quite surprisingly, the inventors have discovered, according to aspects of the invention a class of formulations for delivering mRNA vaccines in vivo that results in significantly enhanced, and in many respects synergistic, immune responses including enhanced antigen generation and functional antibody production with neutralization capability. These results can be achieved even when significantly lower doses of the mRNA are administered in comparison with mRNA doses used in other classes of lipid based formulations. The formulations of the invention have demonstrated significant unexpected in vivo immune responses sufficient to establish the efficacy of functional mRNA vaccines as prophylactic and therapeutic agents. Additionally, self-replicating RNA vaccines rely on viral replication pathways to deliver enough RNA to a cell to produce an immunogenic response. The formulations of the invention do not require viral replication to produce enough protein to result in a strong immune response. Thus, the mRNA of the invention are not self-replicating RNA and do not include components necessary for viral replication.

The invention involves, in some aspects, the surprising finding that lipid nanoparticle (LNP) formulations significantly enhance the effectiveness of mRNA vaccines, including chemically modified and unmodified mRNA vaccines. The efficacy of mRNA vaccines formulated in LNP was examined in vivo using several distinct antigens. The results presented herein demonstrate the unexpected superior efficacy of the mRNA vaccines formulated in LNP over other commercially available vaccines.

In addition to providing an enhanced immune response, the formulations of the invention generate a more rapid immune response with fewer doses of antigen than other vaccines tested. The mRNA-LNP formulations of the invention also produce quantitatively and qualitatively better immune responses than vaccines formulated in a different carriers.

The data described herein demonstrate that the formulations of the invention produced significant unexpected improvements over existing antigen vaccines. Additionally, the mRNA-LNP formulations of the invention are superior to other vaccines even when the dose of mRNA is lower than other vaccines.

The LNP used in the studies described herein has been used previously to deliver siRNA in various animal models as well as in humans. In view of the observations made in association with the siRNA delivery of LNP formulations, the fact that LNP is useful in vaccines is quite surprising. It has been observed that therapeutic delivery of siRNA formulated in LNP causes an undesirable inflammatory response associated with a transient IgM response, typically leading to a reduction in antigen production and a compromised immune response. In contrast to the findings observed with siRNA, the LNP-mRNA formulations of the invention are demonstrated herein to generate enhanced IgG levels, sufficient for prophylactic and therapeutic methods rather than transient IgM responses.

Nucleic Acids/Polynucleotides

Tropical disease vaccines, as provided herein, comprise at least one (one or more) ribonucleic acid (RNA) (e.g., mRNA) polynucleotide having an open reading frame encoding at least one Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide. The term “nucleic acid” includes any compound and/or substance that comprises a polymer of nucleotides (nucleotide monomer). These polymers are referred to as polynucleotides. Thus, the terms “nucleic acid” and “polynucleotide” are used interchangeably.

Nucleic acids may be or may include, for example, ribonucleic acids (RNAs), deoxyribonucleic acids (DNAs), threose nucleic acids (TNAs), glycol nucleic acids (GNAs), peptide nucleic acids (PNAs), locked nucleic acids (LNAs, including LNA having a β-D-ribo configuration, α-LNA having an α-L-ribo configuration (a diastereomer of LNA), 2′-amino-LNA having a 2′-amino functionalization, and 2′-amino-α-LNA having a 2′-amino functionalization), ethylene nucleic acids (ENA), cyclohexenyl nucleic acids (CeNA) or chimeras or combinations thereof.

In some embodiments, polynucleotides of the present disclosure function as messenger RNA (mRNA). “Messenger RNA” (mRNA) refers to any polynucleotide that encodes a (at least one) polypeptide (a naturally-occurring, non-naturally-occurring, or modified polymer of amino acids) and can be translated to produce the encoded polypeptide in vitro, in vivo, in situ or ex vivo. The skilled artisan will appreciate that, except where otherwise noted, polynucleotide sequences set forth in the instant application will recite “T”s in a representative DNA sequence but where the sequence represents RNA (e.g., mRNA), the “T”s would be substituted for “U”s. Thus, any of the RNA polynucleotides encoded by a DNA identified by a particular sequence identification number may also comprise the corresponding RNA (e.g., mRNA) sequence encoded by the DNA, where each “T” of the DNA sequence is substituted with “U.”

The basic components of an mRNA molecule typically include at least one coding region, a 5′ untranslated region (UTR), a 3′ UTR, a 5′ cap and a poly-A tail. Polynucleotides of the present disclosure may function as mRNA but can be distinguished from wild-type mRNA in their functional and/or structural design features, which serve to overcome existing problems of effective polypeptide expression using nucleic-acid based therapeutics.

In some embodiments, a RNA polynucleotide of an RNA (e.g., mRNA) vaccine encodes 2-10, 2-9, 2-8, 2-7, 2-6, 2-5, 2-4, 2-3, 3-10, 3-9, 3-8, 3-7, 3-6, 3-5, 3-4, 4-10, 4-9, 4-8, 4-7, 4-6, 4-5, 5-10, 5-9, 5-8, 5-7, 5-6, 6-10, 6-9, 6-8, 6-7, 7-10, 7-9, 7-8, 8-10, 8-9 or 9-10 antigenic polypeptides. In some embodiments, a RNA (e.g., mRNA) polynucleotide of a tropical disease vaccine encodes at least 10, 20, 30, 40, 50, 60, 70, 80, 90 or 100 antigenic polypeptides. In some embodiments, a RNA (e.g., mRNA) polynucleotide of a tropical disease vaccine encodes at least 100 or at least 200 antigenic polypeptides. In some embodiments, a RNA polynucleotide of a tropical disease vaccine encodes 1-10, 5-15, 10-20, 15-25, 20-30, 25-35, 30-40, 35-45, 40-50, 1-50, 1-100, 2-50 or 2-100 antigenic polypeptides.

Polynucleotides of the present disclosure, in some embodiments, are codon optimized. Codon optimization methods are known in the art and may be used as provided herein. Codon optimization, in some embodiments, may be used to match codon frequencies in target and host organisms to ensure proper folding; bias GC content to increase mRNA stability or reduce secondary structures; minimize tandem repeat codons or base runs that may impair gene construction or expression; customize transcriptional and translational control regions; insert or remove protein trafficking sequences; remove/add post translation modification sites in encoded protein (e.g. glycosylation sites); add, remove or shuffle protein domains; insert or delete restriction sites; modify ribosome binding sites and mRNA degradation sites; adjust translational rates to allow the various domains of the protein to fold properly; or to reduce or eliminate problem secondary structures within the polynucleotide. Codon optimization tools, algorithms and services are known in the art—non-limiting examples include services from GeneArt (Life Technologies), DNA2.0 (Menlo Park Calif.) and/or proprietary methods. In some embodiments, the open reading frame (ORF) sequence is optimized using optimization algorithms.

In some embodiments, a codon optimized sequence shares less than 95% sequence identity, less than 90% sequence identity, less than 85% sequence identity, less than 80% sequence identity, or less than 75% sequence identity to a naturally-occurring or wild-type sequence (e.g., a naturally-occurring or wild-type mRNA sequence encoding a polypeptide or protein of interest (e.g., an antigenic protein or antigenic polypeptide)).

In some embodiments, a codon-optimized sequence shares between 65% and 85% (e.g., between about 67% and about 85%, or between about 67% and about 80%) sequence identity to a naturally-occurring sequence or a wild-type sequence (e.g., a naturally-occurring or wild-type mRNA sequence encoding a polypeptide or protein of interest (e.g., an antigenic protein or polypeptide)). In some embodiments, a codon-optimized sequence shares between 65% and 75%, or about 80% sequence identity to a naturally-occurring sequence or wild-type sequence (e.g., a naturally-occurring or wild-type mRNA sequence encoding a polypeptide or protein of interest (e.g., an antigenic protein or polypeptide)).

In some embodiments a codon-optimized RNA (e.g., mRNA) may, for instance, be one in which the levels of G/C are enhanced. The G/C-content of nucleic acid molecules may influence the stability of the RNA. RNA having an increased amount of guanine (G) and/or cytosine (C) residues may be functionally more stable than nucleic acids containing a large amount of adenine (A) and thymine (T) or uracil (U) nucleotides. WO02/098443 discloses a pharmaceutical composition containing an mRNA stabilized by sequence modifications in the translated region. Due to the degeneracy of the genetic code, the modifications work by substituting existing codons for those that promote greater RNA stability without changing the resulting amino acid. The approach is limited to coding regions of the RNA.

Antigens/Antigenic Polypeptides

In some embodiments, an antigenic polypeptide (e.g., at least one Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide) is longer than 25 amino acids and shorter than 50 amino acids. Polypeptides include gene products, naturally occurring polypeptides, synthetic polypeptides, homologs, orthologs, paralogs, fragments and other equivalents, variants, and analogs of the foregoing. A polypeptide may be a single molecule or may be a multi-molecular complex such as a dimer, trimer or tetramer. Polypeptides may also comprise single chain polypeptides or multichain polypeptides, such as antibodies or insulin, and may be associated or linked to each other. Most commonly, disulfide linkages are found in multichain polypeptides. The term “polypeptide” may also apply to amino acid polymers in which at least one amino acid residue is an artificial chemical analogue of a corresponding naturally-occurring amino acid.

A “polypeptide variant” is a molecule that differs in its amino acid sequence relative to a native sequence or a reference sequence. Amino acid sequence variants may possess substitutions, deletions, insertions, or a combination of any two or three of the foregoing, at certain positions within the amino acid sequence, as compared to a native sequence or a reference sequence. Ordinarily, variants possess at least 50% identity to a native sequence or a reference sequence. In some embodiments, variants share at least 80% identity or at least 90% identity with a native sequence or a reference sequence.

In some embodiments “variant mimics” are provided. A “variant mimic” contains at least one amino acid that would mimic an activated sequence. For example, glutamate may serve as a mimic for phosphoro-threonine and/or phosphoro-serine. Alternatively, variant mimics may result in deactivation or in an inactivated product containing the mimic. For example, phenylalanine may act as an inactivating substitution for tyrosine, or alanine may act as an inactivating substitution for serine.

“Orthologs” refers to genes in different species that evolved from a common ancestral gene by speciation. Normally, orthologs retain the same function in the course of evolution. Identification of orthologs is important for reliable prediction of gene function in newly sequenced genomes.

“Analogs” is meant to include polypeptide variants that differ by one or more amino acid alterations, for example, substitutions, additions or deletions of amino acid residues that still maintain one or more of the properties of the parent or starting polypeptide.

The present disclosure provides several types of compositions that are polynucleotide or polypeptide based, including variants and derivatives. These include, for example, substitutional, insertional, deletion and covalent variants and derivatives. The term “derivative” is synonymous with the term “variant” and generally refers to a molecule that has been modified and/or changed in any way relative to a reference molecule or a starting molecule.

As such, polynucleotides encoding peptides or polypeptides containing substitutions, insertions and/or additions, deletions and covalent modifications with respect to reference sequences, in particular the polypeptide sequences disclosed herein, are included within the scope of this disclosure. For example, sequence tags or amino acids, such as one or more lysines, can be added to peptide sequences (e.g., at the N-terminal or C-terminal ends). Sequence tags can be used for peptide detection, purification or localization. Lysines can be used to increase peptide solubility or to allow for biotinylation. Alternatively, amino acid residues located at the carboxy and amino terminal regions of the amino acid sequence of a peptide or protein may optionally be deleted providing for truncated sequences. Certain amino acids (e.g., C-terminal residues or N-terminal residues) alternatively may be deleted depending on the use of the sequence, as for example, expression of the sequence as part of a larger sequence that is soluble, or linked to a solid support.

“Substitutional variants” when referring to polypeptides are those that have at least one amino acid residue in a native or starting sequence removed and a different amino acid inserted in its place at the same position. Substitutions may be single, where only one amino acid in the molecule has been substituted, or they may be multiple, where two or more (e.g., 3, 4 or 5) amino acids have been substituted in the same molecule.

As used herein the term “conservative amino acid substitution” refers to the substitution of an amino acid that is normally present in the sequence with a different amino acid of similar size, charge, or polarity. Examples of conservative substitutions include the substitution of a non-polar (hydrophobic) residue such as isoleucine, valine and leucine for another non-polar residue. Likewise, examples of conservative substitutions include the substitution of one polar (hydrophilic) residue for another such as between arginine and lysine, between glutamine and asparagine, and between glycine and serine. Additionally, the substitution of a basic residue such as lysine, arginine or histidine for another, or the substitution of one acidic residue such as aspartic acid or glutamic acid for another acidic residue are additional examples of conservative substitutions. Examples of non-conservative substitutions include the substitution of a non-polar (hydrophobic) amino acid residue such as isoleucine, valine, leucine, alanine, methionine for a polar (hydrophilic) residue such as cysteine, glutamine, glutamic acid or lysine and/or a polar residue for a non-polar residue.

“Features” when referring to polypeptide or polynucleotide are defined as distinct amino acid sequence-based or nucleotide-based components of a molecule respectively. Features of the polypeptides encoded by the polynucleotides include surface manifestations, local conformational shape, folds, loops, half-loops, domains, half-domains, sites, termini and any combination(s) thereof.

As used herein when referring to polypeptides the term “domain” refers to a motif of a polypeptide having one or more identifiable structural or functional characteristics or properties (e.g., binding capacity, serving as a site for protein-protein interactions).

As used herein when referring to polypeptides the terms “site” as it pertains to amino acid based embodiments is used synonymously with “amino acid residue” and “amino acid side chain.” As used herein when referring to polynucleotides the terms “site” as it pertains to nucleotide based embodiments is used synonymously with “nucleotide.” A site represents a position within a peptide or polypeptide or polynucleotide that may be modified, manipulated, altered, derivatized or varied within the polypeptide-based or polynucleotide-based molecules.

As used herein the terms “termini” or “terminus” when referring to polypeptides or polynucleotides refer to an extremity of a polypeptide or polynucleotide respectively. Such extremity is not limited only to the first or final site of the polypeptide or polynucleotide but may include additional amino acids or nucleotides in the terminal regions. Polypeptide-based molecules may be characterized as having both an N-terminus (terminated by an amino acid with a free amino group (NH₂)) and a C-terminus (terminated by an amino acid with a free carboxyl group (COOH)). Proteins are in some cases made up of multiple polypeptide chains brought together by disulfide bonds or by non-covalent forces (multimers, oligomers). These proteins have multiple N- and C-termini. Alternatively, the termini of the polypeptides may be modified such that they begin or end, as the case may be, with a non-polypeptide based moiety such as an organic conjugate.

As recognized by those skilled in the art, protein fragments, functional protein domains, and homologous proteins are also considered to be within the scope of polypeptides of interest. For example, provided herein is any protein fragment (meaning a polypeptide sequence at least one amino acid residue shorter than a reference polypeptide sequence but otherwise identical) of a reference protein having a length of 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 or longer than 100 amino acids. In another example, any protein that includes a stretch of 20, 30, 40, 50, or 100 (contiguous) amino acids that are 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 100% identical to any of the sequences described herein can be utilized in accordance with the disclosure. In some embodiments, a polypeptide includes 2, 3, 4, 5, 6, 7, 8, 9, 10, or more mutations as shown in any of the sequences provided herein or referenced herein. In another example, any protein that includes a stretch of 20, 30, 40, 50, or 100 amino acids that are greater than 80%, 90%, 95%, or 100% identical to any of the sequences described herein, wherein the protein has a stretch of 5, 10, 15, 20, 25, or 30 amino acids that are less than 80%, 75%, 70%, 65% to 60% identical to any of the sequences described herein can be utilized in accordance with the disclosure.

Polypeptide or polynucleotide molecules of the present disclosure may share a certain degree of sequence similarity or identity with the reference molecules (e.g., reference polypeptides or reference polynucleotides), for example, with art-described molecules (e.g., engineered or designed molecules or wild-type molecules). The term “identity,” as known in the art, refers to a relationship between the sequences of two or more polypeptides or polynucleotides, as determined by comparing the sequences. In the art, identity also means the degree of sequence relatedness between two sequences as determined by the number of matches between strings of two or more amino acid residues or nucleic acid residues. Identity measures the percent of identical matches between the smaller of two or more sequences with gap alignments (if any) addressed by a particular mathematical model or computer program (e.g., “algorithms”). Identity of related peptides can be readily calculated by known methods. “% identity” as it applies to polypeptide or polynucleotide sequences is defined as the percentage of residues (amino acid residues or nucleic acid residues) in the candidate amino acid or nucleic acid sequence that are identical with the residues in the amino acid sequence or nucleic acid sequence of a second sequence after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent identity. Methods and computer programs for the alignment are well known in the art. Identity depends on a calculation of percent identity but may differ in value due to gaps and penalties introduced in the calculation. Generally, variants of a particular polynucleotide or polypeptide have at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% but less than 100% sequence identity to that particular reference polynucleotide or polypeptide as determined by sequence alignment programs and parameters described herein and known to those skilled in the art. Such tools for alignment include those of the BLAST suite (Stephen F. Altschul, et al. (1997). “Gapped BLAST and PSI-BLAST: a new generation of protein database search programs,” Nucleic Acids Res. 25:3389-3402). Another popular local alignment technique is based on the Smith-Waterman algorithm (Smith, T. F. & Waterman, M. S. (1981) “Identification of common molecular subsequences.” J. Mol. Biol. 147:195-197). A general global alignment technique based on dynamic programming is the Needleman-Wunsch algorithm (Needleman, S. B. & Wunsch, C. D. (1970) “A general method applicable to the search for similarities in the amino acid sequences of two proteins.” J. Mol. Biol. 48:443-453). More recently, a Fast Optimal Global Sequence Alignment Algorithm (FOGSAA) was developed that purportedly produces global alignment of nucleotide and protein sequences faster than other optimal global alignment methods, including the Needleman-Wunsch algorithm. Other tools are described herein, specifically in the definition of “identity” below.

As used herein, the term “homology” refers to the overall relatedness between polymeric molecules, e.g. between nucleic acid molecules (e.g. DNA molecules and/or RNA molecules) and/or between polypeptide molecules. Polymeric molecules (e.g. nucleic acid molecules (e.g. DNA molecules and/or RNA molecules) and/or polypeptide molecules) that share a threshold level of similarity or identity determined by alignment of matching residues are termed homologous. Homology is a qualitative term that describes a relationship between molecules and can be based upon the quantitative similarity or identity. Similarity or identity is a quantitative term that defines the degree of sequence match between two compared sequences. In some embodiments, polymeric molecules are considered to be “homologous” to one another if their sequences are at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% identical or similar. The term “homologous” necessarily refers to a comparison between at least two sequences (polynucleotide or polypeptide sequences). Two polynucleotide sequences are considered homologous if the polypeptides they encode are at least 50%, 60%, 70%, 80%, 90%, 95%, or even 99% for at least one stretch of at least 20 amino acids. In some embodiments, homologous polynucleotide sequences are characterized by the ability to encode a stretch of at least 4-5 uniquely specified amino acids. For polynucleotide sequences less than 60 nucleotides in length, homology is determined by the ability to encode a stretch of at least 4-5 uniquely specified amino acids. Two protein sequences are considered homologous if the proteins are at least 50%, 60%, 70%, 80%, or 90% identical for at least one stretch of at least 20 amino acids.

Homology implies that the compared sequences diverged in evolution from a common origin. The term “homolog” refers to a first amino acid sequence or nucleic acid sequence (e.g., gene (DNA or RNA) or protein sequence) that is related to a second amino acid sequence or nucleic acid sequence by descent from a common ancestral sequence. The term “homolog” may apply to the relationship between genes and/or proteins separated by the event of speciation or to the relationship between genes and/or proteins separated by the event of genetic duplication. “Orthologs” are genes (or proteins) in different species that evolved from a common ancestral gene (or protein) by speciation. Typically, orthologs retain the same function in the course of evolution. “Paralogs” are genes (or proteins) related by duplication within a genome. Orthologs retain the same function in the course of evolution, whereas paralogs evolve new functions, even if these are related to the original one.

The term “identity” refers to the overall relatedness between polymeric molecules, for example, between polynucleotide molecules (e.g. DNA molecules and/or RNA molecules) and/or between polypeptide molecules. Calculation of the percent identity of two polynucleic acid sequences, for example, can be performed by aligning the two sequences for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second nucleic acid sequences for optimal alignment and non-identical sequences can be disregarded for comparison purposes). In certain embodiments, the length of a sequence aligned for comparison purposes is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or 100% of the length of the reference sequence. The nucleotides at corresponding nucleotide positions are then compared. When a position in the first sequence is occupied by the same nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which needs to be introduced for optimal alignment of the two sequences. The comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. For example, the percent identity between two nucleic acid sequences can be determined using methods such as those described in Computational Molecular Biology, Lesk, A. M., ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, D. W., ed., Academic Press, New York, 1993; Sequence Analysis in Molecular Biology, von Heinje, G., Academic Press, 1987; Computer Analysis of Sequence Data, Part I, Griffin, A. M., and Griffin, H. G., eds., Humana Press, New Jersey, 1994; and Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M Stockton Press, New York, 1991; each of which is incorporated herein by reference. For example, the percent identity between two nucleic acid sequences can be determined using the algorithm of Meyers and Miller (CABIOS, 1989, 4:11-17), which has been incorporated into the ALIGN program (version 2.0) using a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4. The percent identity between two nucleic acid sequences can, alternatively, be determined using the GAP program in the GCG software package using an NWSgapdna.CMP matrix. Methods commonly employed to determine percent identity between sequences include, but are not limited to those disclosed in Carillo, H., and Lipman, D., SIAM J Applied Math., 48:1073 (1988); incorporated herein by reference. Techniques for determining identity are codified in publicly available computer programs. Exemplary computer software to determine homology between two sequences include, but are not limited to, GCG program package, Devereux, J., et al., Nucleic Acids Research, 12, 387 (1984)), BLASTP, BLASTN, and FASTA Altschul, S. F. et al., J. Molec. Biol., 215, 403 (1990)).

Multiprotein and Multicomponent Vaccines

The present disclosure encompasses tropical disease vaccines comprising multiple RNA (e.g., mRNA) polynucleotides, each encoding a single antigenic polypeptide, as well as tropical disease vaccines comprising a single RNA polynucleotide encoding more than one antigenic polypeptide (e.g., as a fusion polypeptide). Thus, a vaccine composition comprising a RNA (e.g., mRNA) polynucleotide having an open reading frame encoding a first antigenic polypeptide and a RNA (e.g., mRNA) polynucleotide having an open reading frame encoding a second antigenic polypeptide encompasses (a) vaccines that comprise a first RNA polynucleotide encoding a first antigenic polypeptide and a second RNA polynucleotide encoding a second antigenic polypeptide, and (b) vaccines that comprise a single RNA polynucleotide encoding a first and second antigenic polypeptide (e.g., as a fusion polypeptide). RNA (e.g., mRNA) vaccines of the present disclosure, in some embodiments, comprise 2-10 (e.g., 2, 3, 4, 5, 6, 7, 8, 9 or 10), or more, RNA polynucleotides having an open reading frame, each of which encodes a different antigenic polypeptide (or a single RNA polynucleotide encoding 2-10, or more, different antigenic polypeptides). The antigenic polypeptides may be selected from Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and YFV antigenic polypeptides.

In some embodiments, a tropical disease vaccine comprises a RNA (e.g., mRNA) polynucleotide having an open reading frame encoding a viral capsid protein, a RNA (e.g., mRNA) polynucleotide having an open reading frame encoding a viral premembrane/membrane protein, and a RNA (e.g., mRNA) polynucleotide having an open reading frame encoding a viral envelope protein. In some embodiments, a tropical disease vaccine comprises a RNA (e.g., mRNA) polynucleotide having an open reading frame encoding a viral fusion (F) protein and a RNA polynucleotide having an open reading frame encoding a viral major surface glycoprotein (G protein). In some embodiments, a vaccine comprises a RNA (e.g., mRNA) polynucleotide having an open reading frame encoding a viral F protein. In some embodiments, a vaccine comprises a RNA (e.g., mRNA) polynucleotide having an open reading frame encoding a viral G protein. In some embodiments, a vaccine comprises a RNA (e.g., mRNA) polynucleotide having an open reading frame encoding a HN protein.

In some embodiments, a multicomponent vaccine comprises at least one RNA (e.g., mRNA) polynucleotide encoding at least one antigenic polypeptide fused to a signal peptide (e.g., SEQ ID NO: 304-307). The signal peptide may be fused at the N-terminus or the C-terminus of an antigenic polypeptide. An antigenic polypeptide fused to a signal peptide may be selected from Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and YFV antigenic polypeptides.

Signal Peptides

In some embodiments, antigenic polypeptides encoded by tropical disease RNA (e.g., mRNA) polynucleotides comprise a signal peptide. Signal peptides, comprising the N-terminal 15-60 amino acids of proteins, are typically needed for the translocation across the membrane on the secretory pathway and, thus, universally control the entry of most proteins both in eukaryotes and prokaryotes to the secretory pathway. Signal peptides generally include three regions: an N-terminal region of differing length, which usually comprises positively charged amino acids; a hydrophobic region; and a short carboxy-terminal peptide region. In eukaryotes, the signal peptide of a nascent precursor protein (pre-protein) directs the ribosome to the rough endoplasmic reticulum (ER) membrane and initiates the transport of the growing peptide chain across it for processing. ER processing produces mature proteins, wherein the signal peptide is cleaved from precursor proteins, typically by a ER-resident signal peptidase of the host cell, or they remain uncleaved and function as a membrane anchor. A signal peptide may also facilitate the targeting of the protein to the cell membrane. The signal peptide, however, is not responsible for the final destination of the mature protein. Secretory proteins devoid of additional address tags in their sequence are by default secreted to the external environment. During recent years, a more advanced view of signal peptides has evolved, showing that the functions and immunodominance of certain signal peptides are much more versatile than previously anticipated.

Tropical disease vaccines of the present disclosure may comprise, for example, RNA (e.g., mRNA) polynucleotides encoding an artificial signal peptide, wherein the signal peptide coding sequence is operably linked to and is in frame with the coding sequence of the antigenic polypeptide. Thus, tropical disease vaccines of the present disclosure, in some embodiments, produce an antigenic polypeptide (e.g., a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide) fused to a signal peptide. In some embodiments, a signal peptide is fused to the N-terminus of the antigenic polypeptide. In some embodiments, a signal peptide is fused to the C-terminus of the antigenic polypeptide.

In some embodiments, the signal peptide fused to the antigenic polypeptide is an artificial signal peptide. In some embodiments, an artificial signal peptide fused to the antigenic polypeptide encoded by the RNA (e.g., mRNA) vaccine is obtained from an immunoglobulin protein, e.g., an IgE signal peptide or an IgG signal peptide. In some embodiments, a signal peptide fused to the antigenic polypeptide encoded by a RNA (e.g., mRNA) vaccine is an Ig heavy chain epsilon-1 signal peptide (IgE HC SP) having the sequence of: MDWTWILFLVAAATRVHS; SEQ ID NO: 424. In some embodiments, a signal peptide fused to the antigenic polypeptide encoded by the (e.g., mRNA) RNA (e.g., mRNA) vaccine is an IgGk chain V-III region HAH signal peptide (IgGk SP) having the sequence of METPAQLLFLLLLWLPDTTG; SEQ ID NO: 423. In some embodiments, the signal peptide is selected from: Japanese encephalitis PRM signal sequence (MLGSNSGQRVVFTILLLLVAPAYS; SEQ ID NO: 425), VSINVg protein signal sequence (MKCLLYLAFLFIGVNCA; SEQ ID NO: 426) and Japanese encephalitis JEV signal sequence (MWLVSLAIVTACAGA; SEQ ID NO: 427).

In some embodiments, the antigenic polypeptide encoded by a RNA (e.g., mRNA) vaccine comprises an amino acid sequence identified by any one of SEQ ID NO: 13-17, 22-29, 44-47, 52-54, 59-64, 97-117, 156-222, 469, 259-291 or 402-413 fused to a signal peptide identified by any one of SEQ ID NO: 423-427. The examples disclosed herein are not meant to be limiting and any signal peptide that is known in the art to facilitate targeting of a protein to ER for processing and/or targeting of a protein to the cell membrane may be used in accordance with the present disclosure.

A signal peptide may have a length of 15-60 amino acids. For example, a signal peptide may have a length of 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60 amino acids. In some embodiments, a signal peptide has a length of 20-60, 25-60, 30-60, 35-60, 40-60, 45-60, 50-60, 55-60, 15-55, 20-55, 25-55, 30-55, 35-55, 40-55, 45-55, 50-55, 15-50, 20-50, 25-50, 30-50, 35-50, 40-50, 45-50, 15-45, 20-45, 25-45, 30-45, 35-45, 40-45, 15-40, 20-40, 25-40, 30-40, 35-40, 15-35, 20-35, 25-35, 30-35, 15-30, 20-30, 25-30, 15-25, 20-25, or 15-20 amino acids.

A signal peptide is typically cleaved from the nascent polypeptide at the cleavage junction during ER processing. The mature antigenic polypeptide produce by a tropical disease RNA (e.g., mRNA) vaccine of the present disclosure typically does not comprise a signal peptide.

Chemical Modifications

Tropical disease vaccines of the present disclosure, in some embodiments, comprise at least RNA (e.g. mRNA) polynucleotide having an open reading frame encoding at least one antigenic polypeptide that comprises at least one chemical modification.

The terms “chemical modification” and “chemically modified” refer to modification with respect to adenosine (A), guanosine (G), uridine (U), thymidine (T) or cytidine (C) ribonucleosides or deoxyribnucleosides in at least one of their position, pattern, percent or population. Generally, these terms do not refer to the ribonucleotide modifications in naturally occurring 5′-terminal mRNA cap moieties. With respect to a polypeptide, the term “modification” refers to a modification relative to the canonical set 20 amino acids. Polypeptides, as provided herein, are also considered “modified” of they contain amino acid substitutions, insertions or a combination of substitutions and insertions.

Polynucleotides (e.g., RNA polynucleotides, such as mRNA polynucleotides), in some embodiments, comprise various (more than one) different modifications. In some embodiments, a particular region of a polynucleotide contains one, two or more (optionally different) nucleoside or nucleotide modifications. In some embodiments, a modified RNA polynucleotide (e.g., a modified mRNA polynucleotide), introduced to a cell or organism, exhibits reduced degradation in the cell or organism, respectively, relative to an unmodified polynucleotide. In some embodiments, a modified RNA polynucleotide (e.g., a modified mRNA polynucleotide), introduced into a cell or organism, may exhibit reduced immunogenicity in the cell or organism, respectively (e.g., a reduced innate response).

Modifications of polynucleotides include, without limitation, those described herein. Polynucleotides (e.g., RNA polynucleotides, such as mRNA polynucleotides) may comprise modifications that are naturally-occurring, non-naturally-occurring or the polynucleotide may comprise a combination of naturally-occurring and non-naturally-occurring modifications. Polynucleotides may include any useful modification, for example, of a sugar, a nucleobase, or an internucleoside linkage (e.g., to a linking phosphate, to a phosphodiester linkage or to the phosphodiester backbone).

Polynucleotides (e.g., RNA polynucleotides, such as mRNA polynucleotides), in some embodiments, comprise non-natural modified nucleotides that are introduced during synthesis or post-synthesis of the polynucleotides to achieve desired functions or properties. The modifications may be present on an internucleotide linkages, purine or pyrimidine bases, or sugars. The modification may be introduced with chemical synthesis or with a polymerase enzyme at the terminal of a chain or anywhere else in the chain. Any of the regions of a polynucleotide may be chemically modified.

The present disclosure provides for modified nucleosides and nucleotides of a polynucleotide (e.g., RNA polynucleotides, such as mRNA polynucleotides). A “nucleoside” refers to a compound containing a sugar molecule (e.g., a pentose or ribose) or a derivative thereof in combination with an organic base (e.g., a purine or pyrimidine) or a derivative thereof (also referred to herein as “nucleobase”). A “nucleotide” refers to a nucleoside, including a phosphate group. Modified nucleotides may by synthesized by any useful method, such as, for example, chemically, enzymatically, or recombinantly, to include one or more modified or non-natural nucleosides. Polynucleotides may comprise a region or regions of linked nucleosides. Such regions may have variable backbone linkages. The linkages may be standard phosphodiester linkages, in which case the polynucleotides would comprise regions of nucleotides.

Modified nucleotide base pairing encompasses not only the standard adenosine-thymine, adenosine-uracil, or guanosine-cytosine base pairs, but also base pairs formed between nucleotides and/or modified nucleotides comprising non-standard or modified bases, wherein the arrangement of hydrogen bond donors and hydrogen bond acceptors permits hydrogen bonding between a non-standard base and a standard base or between two complementary non-standard base structures. One example of such non-standard base pairing is the base pairing between the modified nucleotide inosine and adenine, cytosine or uracil. Any combination of base/sugar or linker may be incorporated into polynucleotides of the present disclosure.

Modifications of polynucleotides (e.g., RNA polynucleotides, such as mRNA polynucleotides) that are useful in the vaccines of the present disclosure include, but are not limited to the following: 2-methylthio-N6-(cis-hydroxyisopentenyl)adenosine; 2-methylthio-N6-methyladenosine; 2-methylthio-N6-threonyl carbamoyladenosine; N6-glycinylcarbamoyladenosine; N6-isopentenyladenosine; N6-methyladenosine; N6-threonylcarbamoyladeno sine; 1,2′-O-dimethyladenosine; 1-methyladenosine; 2′-O-methyladenosine; 2′-O-ribosyladenosine (phosphate); 2-methyladenosine; 2-methylthio-N6 isopentenyladenosine; 2-methylthio-N6-hydroxynorvalyl carbamoyladenosine; 2′-O-methyladenosine; 2′-O-ribosyladenosine (phosphate); Isopentenyladenosine; N6-(cis-hydroxyisopentenyl)adenosine; N6,2′-O-dimethyladenosine; N6,2′-O-dimethyladenosine; N6,N6,2′-O-trimethyladenosine; N6,N6-dimethyladenosine; N6-acetyladenosine; N6-hydroxynorvalylcarbamoyladenosine; N6-methyl-N6-threonylcarbamoyladenosine; 2-methyladenosine; 2-methylthio-N6-isopentenyladenosine; 7-deaza-adenosine; N1-methyl-adenosine; N6, N6 (dimethyl)adenine; N6-cis-hydroxy-isopentenyl-adenosine; α-thio-adenosine; 2 (amino)adenine; 2 (aminopropyl)adenine; 2 (methylthio) N6 (isopentenyl)adenine; 2-(alkyl)adenine; 2-(aminoalkyl)adenine; 2-(aminopropyl)adenine; 2-(halo)adenine; 2-(halo)adenine; 2-(propyl)adenine; 2′-Amino-2′-deoxy-ATP; 2′-Azido-2′-deoxy-ATP; 2′-Deoxy-2′-a-aminoadenosine TP; 2′-Deoxy-2′-a-azidoadenosine TP; 6 (alkyl)adenine; 6 (methyl)adenine; 6-(alkyl)adenine; 6-(methyl)adenine; 7 (deaza)adenine; 8 (alkenyl)adenine; 8 (alkynyl)adenine; 8 (amino)adenine; 8 (thioalkyl)adenine; 8-(alkenyl)adenine; 8-(alkyl)adenine; 8-(alkynyl)adenine; 8-(amino)adenine; 8-(halo)adenine; 8-(hydroxyl)adenine; 8-(thioalkyl)adenine; 8-(thiol)adenine; 8-azido-adeno sine; aza adenine; deaza adenine; N6 (methyl)adenine; N6-(isopentyl)adenine; 7-deaza-8-aza-adenosine; 7-methyladenine; 1-Deazaadenosine TP; 2′Fluoro-N6-Bz-deoxyadenosine TP; 2′-OMe-2-Amino-ATP; 2′O-methyl-N6-Bz-deoxyadenosine TP; 2′-a-Ethynyladenosine TP; 2-aminoadenine; 2-Aminoadenosine TP; 2-Amino-ATP; 2′-a-Trifluoromethyladenosine TP; 2-Azidoadenosine TP; 2′-b-Ethynyladenosine TP; 2-Bromoadenosine TP; 2′-b-Trifluoromethyladenosine TP; 2-Chloroadenosine TP; 2′-Deoxy-2′,2′-difluoroadenosine TP; 2′-Deoxy-2′-a-mercaptoadenosine TP; 2′-Deoxy-2′-a-thiomethoxyadenosine TP; 2′-Deoxy-2′-b-aminoadenosine TP; 2′-Deoxy-2′-b-azidoadenosine TP; 2′-Deoxy-2′-b-bromoadenosine TP; 2′-Deoxy-2′-b-chloroadenosine TP; 2′-Deoxy-2′-b-fluoroadenosine TP; 2′-Deoxy-2′-b-iodoadenosine TP; 2′-Deoxy-2′-b-mercaptoadenosine TP; 2′-Deoxy-2′-b-thiomethoxyadenosine TP; 2-Fluoroadenosine TP; 2-Iodoadenosine TP; 2-Mercaptoadenosine TP; 2-methoxy-adenine; 2-methylthio-adenine; 2-Trifluoromethyladenosine TP; 3-Deaza-3-bromoadenosine TP; 3-Deaza-3-chloroadenosine TP; 3-Deaza-3-fluoroadenosine TP; 3-Deaza-3-iodoadenosine TP; 3-Deazaadenosine TP; 4′-Azidoadenosine TP; 4′-Carbocyclic adenosine TP; 4′-Ethynyladenosine TP; 5′-Homo-adenosine TP; 8-Aza-ATP; 8-bromo-adenosine TP; 8-Trifluoromethyladenosine TP; 9-Deazaadenosine TP; 2-aminopurine; 7-deaza-2,6-diaminopurine; 7-deaza-8-aza-2,6-diaminopurine; 7-deaza-8-aza-2-aminopurine; 2,6-diaminopurine; 7-deaza-8-aza-adenine, 7-deaza-2-aminopurine; 2-thiocytidine; 3-methylcytidine; 5-formylcytidine; 5-hydroxymethylcytidine; 5-methylcytidine; N4-acetylcytidine; 2′-O-methylcytidine; 2′-O-methylcytidine; 5,2′-O-dimethylcytidine; 5-formyl-2′-O-methylcytidine; Lysidine; N4,2′-O-dimethylcytidine; N4-acetyl-2′-O-methylcytidine; N4-methylcytidine; N4,N4-Dimethyl-2′-OMe-Cytidine TP; 4-methylcytidine; 5-aza-cytidine; Pseudo-iso-cytidine; pyrrolo-cytidine; α-thio-cytidine; 2-(thio)cytosine; 2′-Amino-2′-deoxy-CTP; 2′-Azido-2′-deoxy-CTP; 2′-Deoxy-2′-a-aminocytidine TP; 2′-Deoxy-2′-a-azidocytidine TP; 3 (deaza) 5 (aza)cytosine; 3 (methyl)cytosine; 3-(alkyl)cytosine; 3-(deaza) 5 (aza)cytosine; 3-(methyl)cytidine; 4,2′-O-dimethylcytidine; 5 (halo)cytosine; 5 (methyl)cytosine; 5 (propynyl)cytosine; 5 (trifluoromethyl)cytosine; 5-(alkyl)cytosine; 5-(alkynyl)cytosine; 5-(halo)cytosine; 5-(propynyl)cytosine; 5-(trifluoromethyl)cytosine; 5-bromo-cytidine; 5-iodo-cytidine; 5-propynyl cytosine; 6-(azo)cytosine; 6-aza-cytidine; aza cytosine; deaza cytosine; N4 (acetyl)cytosine; 1-methyl-1-deaza-pseudoisocytidine; 1-methyl-pseudoisocytidine; 2-methoxy-5-methyl-cytidine; 2-methoxy-cytidine; 2-thio-5-methyl-cytidine; 4-methoxy-1-methyl-pseudoisocytidine; 4-methoxy-pseudoisocytidine; 4-thio-1-methyl-1-deaza-pseudoisocytidine; 4-thio-1-methyl-pseudoisocytidine; 4-thio-pseudoisocytidine; 5-aza-zebularine; 5-methyl-zebularine; pyrrolo-pseudoisocytidine; Zebularine; (E)-5-(2-Bromo-vinyl)cytidine TP; 2,2′-anhydro-cytidine TP hydrochloride; 2′Fluor-N4-Bz-cytidine TP; 2′Fluoro-N4-Acetyl-cytidine TP; 2′-O-Methyl-N4-Acetyl-cytidine TP; 2′O-methyl-N4-Bz-cytidine TP; 2′-a-Ethynylcytidine TP; 2′-a-Trifluoromethylcytidine TP; 2′-b-Ethynylcytidine TP; 2′-b-Trifluoromethylcytidine TP; 2′-Deoxy-2′,2′-difluorocytidine TP; 2′-Deoxy-2′-a-mercaptocytidine TP; 2′-Deoxy-2′-a-thiomethoxycytidine TP; 2′-Deoxy-2′-b-aminocytidine TP; 2′-Deoxy-2′-b-azidocytidine TP; 2′-Deoxy-2′-b-bromocytidine TP; 2′-Deoxy-2′-b-chlorocytidine TP; 2′-Deoxy-2′-b-fluorocytidine TP; 2′-Deoxy-2′-b-iodocytidine TP; 2′-Deoxy-2′-b-mercaptocytidine TP; 2′-Deoxy-2′-b-thiomethoxycytidine TP; 2′-O-Methyl-5-(1-propynyl)cytidine TP; 3′-Ethynylcytidine TP; 4′-Azidocytidine TP; 4′-Carbocyclic cytidine TP; 4′-Ethynylcytidine TP; 5-(1-Propynyl)ara-cytidine TP; 5-(2-Chloro-phenyl)-2-thiocytidine TP; 5-(4-Amino-phenyl)-2-thiocytidine TP; 5-Aminoallyl-CTP; 5-Cyanocytidine TP; 5-Ethynylara-cytidine TP; 5-Ethynylcytidine TP; 5′-Homo-cytidine TP; 5-Methoxycytidine TP; 5-Trifluoromethyl-Cytidine TP; N4-Amino-cytidine TP; N4-Benzoyl-cytidine TP; Pseudoisocytidine; 7-methylguanosine; N2,2′-O-dimethylguanosine; N2-methylguanosine; Wyosine; 1,2′-O-dimethylguanosine; 1-methylguanosine; 2′-O-methylguanosine; 2′-O-ribosylguanosine (phosphate); 2′-O-methylguanosine; 2′-O-ribosylguanosine (phosphate); 7-aminomethyl-7-deazaguanosine; 7-cyano-7-deazaguanosine; Archaeosine; Methylwyo sine; N2,7-dimethylguanosine; N2,N2,2′-O-trimethylguanosine; N2,N2,7-trimethylguanosine; N2,N2-dimethylguanosine; N2,7,2′-O-trimethylguanosine; 6-thio-guanosine; 7-deaza-guanosine; 8-oxo-guanosine; N1-methyl-guanosine; α-thio-guanosine; 2 (propyl)guanine; 2-(alkyl)guanine; 2′-Amino-2′-deoxy-GTP; 2′-Azido-2′-deoxy-GTP; 2′-Deoxy-2′-a-aminoguanosine TP; 2′-Deoxy-2′-a-azidoguanosine TP; 6 (methyl)guanine; 6-(alkyl)guanine; 6-(methyl)guanine; 6-methyl-guanosine; 7 (alkyl)guanine; 7 (deaza)guanine; 7 (methyl)guanine; 7-(alkyl)guanine; 7-(deaza)guanine; 7-(methyl)guanine; 8 (alkyl)guanine; 8 (alkynyl)guanine; 8 (halo)guanine; 8 (thioalkyl)guanine; 8-(alkenyl)guanine; 8-(alkyl)guanine; 8-(alkynyl)guanine; 8-(amino)guanine; 8-(halo)guanine; 8-(hydroxyl)guanine; 8-(thioalkyl)guanine; 8-(thiol)guanine; aza guanine; deaza guanine; N (methyl)guanine; N-(methyl)guanine; 1-methyl-6-thio-guanosine; 6-methoxy-guanosine; 6-thio-7-deaza-8-aza-guanosine; 6-thio-7-deaza-guanosine; 6-thio-7-methyl-guanosine; 7-deaza-8-aza-guanosine; 7-methyl-8-oxo-guanosine; N2,N2-dimethyl-6-thio-guanosine; N2-methyl-6-thio-guanosine; 1-Me-GTP; 2′Fluoro-N2-isobutyl-guanosine TP; 2′O-methyl-N2-isobutyl-guanosine TP; 2′-a-Ethynylguanosine TP; 2′-a-Trifluoromethylguanosine TP; 2′-b-Ethynylguano sine TP; 2′-b-Trifluoromethylguanosine TP; 2′-Deoxy-2′,2′-difluoroguanosine TP; 2′-Deoxy-2′-a-mercaptoguanosine TP; 2′-Deoxy-2′-a-thiomethoxyguanosine TP; 2′-Deoxy-2′-b-aminoguanosine TP; 2′-Deoxy-2′-b-azidoguanosine TP; 2′-Deoxy-2′-b-bromoguanosine TP; 2′-Deoxy-2′-b-chloroguanosine TP; 2′-Deoxy-2′-b-fluoroguanosine TP; 2′-Deoxy-2′-b-iodoguanosine TP; 2′-Deoxy-2′-b-mercaptoguanosine TP; 2′-Deoxy-2′-b-thiomethoxyguanosine TP; 4′-Azidoguanosine TP; 4′-Carbocyclic guanosine TP; 4′-Ethynylguanosine TP; 5′-Homo-guanosine TP; 8-bromo-guanosine TP; 9-Deazaguanosine TP; N2-isobutyl-guanosine TP; 1-methylinosine; Inosine; 1,2′-O-dimethylinosine; 2′-O-methylinosine; 7-methylinosine; 2′-O-methylinosine; Epoxyqueuosine; galactosyl-queuosine; Mannosylqueuosine; Queuosine; allyamino-thymidine; aza thymidine; deaza thymidine; deoxy-thymidine; 2′-O-methyluridine; 2-thiouridine; 3-methyluridine; 5-carboxymethyluridine; 5-hydroxyuridine; 5-methyluridine; 5-taurinomethyl-2-thiouridine; 5-taurinomethyluridine; Dihydrouridine; Pseudouridine; (3-(3-amino-3-carboxypropyl)uridine; 1-methyl-3-(3-amino-5-carboxypropyl)pseudouridine; 1-methylpseduouridine; 1-methyl-pseudouridine; 2′-O-methyluridine; 2′-O-methylpseudouridine; 2′-O-methyluridine; 2-thio-2′-O-methyluridine; 3-(3-amino-3-carboxypropyl)uridine; 3,2′-O-dimethyluridine; 3-Methyl-pseudo-Uridine TP; 4-thiouridine; 5-(carboxyhydroxymethyl)uridine; 5-(carboxyhydroxymethyl)uridine methyl ester; 5,2′-O-dimethyluridine; 5,6-dihydro-uridine; 5-aminomethyl-2-thiouridine; 5-carbamoylmethyl-2′-O-methyluridine; 5-carbamoylmethyluridine; 5-carboxyhydroxymethyluridine; 5-carboxyhydroxymethyluridine methyl ester; 5-carboxymethylaminomethyl-2′-O-methyluridine; 5-carboxymethylaminomethyl-2-thiouridine; 5-carboxymethylaminomethyl-2-thiouridine; 5-carboxymethylaminomethyluridine; 5-carboxymethylaminomethyluridine; 5-Carbamoylmethyluridine TP; 5-methoxycarbonylmethyl-2′-O-methyluridine; 5-methoxycarbonylmethyl-2-thiouridine; 5-methoxycarbonylmethyluridine; 5-methoxyuridine; 5-methyl-2-thiouridine; 5-methylaminomethyl-2-selenouridine; 5-methylaminomethyl-2-thiouridine; 5-methylaminomethyluridine; 5-Methyldihydrouridine; 5-Oxyacetic acid-Uridine TP; 5-Oxyacetic acid-methyl ester-Uridine TP; N1-methyl-pseudo-uridine; uridine 5-oxyacetic acid; uridine 5-oxyacetic acid methyl ester; 3-(3-Amino-3-carboxypropyl)-Uridine TP; 5-(iso-Pentenylaminomethyl)-2-thiouridine TP; 5-(iso-Pentenylaminomethyl)-2′-O-methyluridine TP; 5-(iso-Pentenylaminomethyl)uridine TP; 5-propynyl uracil; α-thio-uridine; 1 (aminoalkylamino-carbonylethylenyl)-2(thio)-pseudouracil; 1 (aminoalkylaminocarbonylethylenyl)-2,4-(dithio)pseudouracil; 1 (aminoalkylaminocarbonylethylenyl)-4 (thio)pseudouracil; 1 (aminoalkylaminocarbonylethylenyl)-pseudouracil; 1 (aminocarbonylethylenyl)-2 (thio)-pseudouracil; 1 (aminocarbonylethylenyl)-2,4-(dithio)pseudouracil; 1 (aminocarbonylethylenyl)-4 (thio)pseudouracil; 1 (aminocarbonylethylenyl)-pseudouracil; 1 substituted 2(thio)-pseudouracil; 1 substituted 2,4-(dithio)pseudouracil; 1 substituted 4 (thio)pseudouracil; 1 substituted pseudouracil; 1-(aminoalkylamino-carbonylethylenyl)-2-(thio)-pseudouracil; 1-Methyl-3-(3-amino-3-carboxypropyl) pseudouridine TP; 1-Methyl-3-(3-amino-3-carboxypropyl)pseudo-UTP; 1-Methyl-pseudo-UTP; 2 (thio)pseudouracil; 2′ deoxy uridine; 2′ fluorouridine; 2-(thio)uracil; 2,4-(dithio)psuedouracil; 2′ methyl, 2′amino, 2′azido, 2′fluro-guanosine; 2′-Amino-2′-deoxy-UTP; 2′-Azido-2′-deoxy-UTP; 2′-Azido-deoxyuridine TP; 2′-O-methylpseudouridine; 2′ deoxy uridine; 2′ fluorouridine; 2′-Deoxy-2′-a-aminouridine TP; 2′-Deoxy-2′-a-azidouridine TP; 2-methylpseudouridine; 3 (3 amino-3 carboxypropyl)uracil; 4 (thio)pseudouracil; 4-(thio)pseudouracil; 4-(thio)uracil; 4-thiouracil; 5 (1,3-diazole-1-alkyl)uracil; 5 (2-aminopropyl)uracil; 5 (aminoalkyl)uracil; 5 (dimethylaminoalkyl)uracil; 5 (guanidiniumalkyl)uracil; 5 (methoxycarbonylmethyl)-2-(thio)uracil; 5 (methoxycarbonyl-methyl)uracil; 5 (methyl) 2 (thio)uracil; 5 (methyl) 2,4 (dithio)uracil; 5 (methyl) 4 (thio)uracil; 5 (methylaminomethyl)-2 (thio)uracil; 5 (methylaminomethyl)-2,4 (dithio)uracil; 5 (methylaminomethyl)-4 (thio)uracil; 5 (propynyl)uracil; 5 (trifluoromethyl)uracil; 5-(2-aminopropyl)uracil; 5-(alkyl)-2-(thio)pseudouracil; 5-(alkyl)-2,4 (dithio)pseudouracil; 5-(alkyl)-4 (thio)pseudouracil; 5-(alkyl)pseudouracil; 5-(alkyl)uracil; 5-(alkynyl)uracil; 5-(allylamino)uracil; 5-(cyanoalkyl)uracil; 5-(dialkylaminoalkyl)uracil; 5-(dimethylaminoalkyl)uracil; 5-(guanidiniumalkyl)uracil; 5-(halo)uracil; 5-(1,3-diazole-1-alkyl)uracil; 5-(methoxy)uracil; 5-(methoxycarbonylmethyl)-2-(thio)uracil; 5-(methoxycarbonyl-methyl)uracil; 5-(methyl) 2(thio)uracil; 5-(methyl) 2,4 (dithio)uracil; 5-(methyl) 4 (thio)uracil; 5-(methyl)-2-(thio)pseudouracil; 5-(methyl)-2,4 (dithio)pseudouracil; 5-(methyl)-4 (thio)pseudouracil; 5-(methyl)pseudouracil; 5-(methylaminomethyl)-2 (thio)uracil; 5-(methylaminomethyl)-2,4(dithio)uracil; 5-(methylaminomethyl)-4-(thio)uracil; 5-(propynyl)uracil; 5-(trifluoromethyl)uracil; 5-aminoallyl-uridine; 5-bromo-uridine; 5-iodo-uridine; 5-uracil; 6 (azo)uracil; 6-(azo)uracil; 6-aza-uridine; allyamino-uracil; aza uracil; deaza uracil; N3 (methyl)uracil; Pseudo-UTP-1-2-ethanoic acid; Pseudouracil; 4-Thio-pseudo-UTP; 1-carboxymethyl-pseudouridine; 1-methyl-1-deaza-pseudouridine; 1-propynyl-uridine; 1-taurinomethyl-1-methyl-uridine; 1-taurinomethyl-4-thio-uridine; 1-taurinomethyl-pseudouridine; 2-methoxy-4-thio-pseudouridine; 2-thio-1-methyl-1-deaza-pseudouridine; 2-thio-1-methyl-pseudouridine; 2-thio-5-aza-uridine; 2-thio-dihydropseudouridine; 2-thio-dihydrouridine; 2-thio-pseudouridine; 4-methoxy-2-thio-pseudouridine; 4-methoxy-pseudouridine; 4-thio-1-methyl-pseudouridine; 4-thio-pseudouridine; 5-aza-uridine; Dihydropseudouridine; (±)1-(2-Hydroxypropyl)pseudouridine TP; (2R)-1-(2-Hydroxypropyl)pseudouridine TP; (2S)-1-(2-Hydroxypropyl)pseudouridine TP; (E)-5-(2-Bromo-vinyl)ara-uridine TP; (E)-5-(2-Bromo-vinyl)uridine TP; (Z)-5-(2-Bromo-vinyl)ara-uridine TP; (Z)-5-(2-Bromo-vinyl)uridine TP; 1-(2,2,2-Trifluoroethyl)-pseudo-UTP; 1-(2,2,3,3,3-Pentafluoropropyl)pseudouridine TP; 1-(2,2-Diethoxyethyl)pseudouridine TP; 1-(2,4,6-Trimethylbenzyl)pseudouridine TP; 1-(2,4,6-Trimethyl-benzyl)pseudo-UTP; 1-(2,4,6-Trimethyl-phenyl)pseudo-UTP; 1-(2-Amino-2-carboxyethyl)pseudo-UTP; 1-(2-Amino-ethyl)pseudo-UTP; 1-(2-Hydroxyethyl)pseudouridine TP; 1-(2-Methoxyethyl)pseudouridine TP; 1-(3,4-Bis-trifluoromethoxybenzyl)pseudouridine TP; 1-(3,4-Dimethoxybenzyl)pseudouridine TP; 1-(3-Amino-3-carboxypropyl)pseudo-UTP; 1-(3-Amino-propyl)pseudo-UTP; 1-(3-Cyclopropyl-prop-2-ynyl)pseudouridine TP; 1-(4-Amino-4-carboxybutyl)pseudo-UTP; 1-(4-Amino-benzyl)pseudo-UTP; 1-(4-Amino-butyl)pseudo-UTP; 1-(4-Amino-phenyl)pseudo-UTP; 1-(4-Azidobenzyl)pseudouridine TP; 1-(4-Bromobenzyl)pseudouridine TP; 1-(4-Chlorobenzyl)pseudouridine TP; 1-(4-Fluorobenzyl)pseudouridine TP; 1-(4-Iodobenzyl)pseudouridine TP; 1-(4-Methanesulfonylbenzyl)pseudouridine TP; 1-(4-Methoxybenzyl)pseudouridine TP; 1-(4-Methoxy-benzyl)pseudo-UTP; 1-(4-Methoxy-phenyl)pseudo-UTP; 1-(4-Methylbenzyl)pseudouridine TP; 1-(4-Methyl-benzyl)pseudo-UTP; 1-(4-Nitrobenzyl)pseudouridine TP; 1-(4-Nitro-benzyl)pseudo-UTP; 1(4-Nitro-phenyl)pseudo-UTP; 1-(4-Thiomethoxybenzyl)pseudouridine TP; 1-(4-Trifluoromethoxybenzyl)pseudouridine TP; 1-(4-Trifluoromethylbenzyl)pseudouridine TP; 1-(5-Amino-pentyl)pseudo-UTP; 1-(6-Amino-hexyl)pseudo-UTP; 1,6-Dimethyl-pseudo-UTP; 1-[3-(2-{2-[2-(2-Aminoethoxy)-ethoxy]-ethoxy}-ethoxy)-propionyl]pseudouridine TP; 1-{3-[2-(2-Aminoethoxy)-ethoxy]-propionyl} pseudouridine TP; 1-Acetylpseudouridine TP; 1-Alkyl-6-(1-propynyl)-pseudo-UTP; 1-Alkyl-6-(2-propynyl)-pseudo-UTP; 1-Alkyl-6-allyl-pseudo-UTP; 1-Alkyl-6-ethynyl-pseudo-UTP; 1-Alkyl-6-homoallyl-pseudo-UTP; 1-Alkyl-6-vinyl-pseudo-UTP; 1-Allylpseudouridine TP; 1-Aminomethyl-pseudo-UTP; 1-Benzoylpseudouridine TP; 1-Benzyloxymethylpseudouridine TP; 1-Benzyl-pseudo-UTP; 1-Biotinyl-PEG2-pseudouridine TP; 1-Biotinylpseudouridine TP; 1-Butyl-pseudo-UTP; 1-Cyanomethylpseudouridine TP; 1-Cyclobutylmethyl-pseudo-UTP; 1-Cyclobutyl-pseudo-UTP; 1-Cycloheptylmethyl-pseudo-UTP; 1-Cycloheptyl-pseudo-UTP; 1-Cyclohexylmethyl-pseudo-UTP; 1-Cyclohexyl-pseudo-UTP; 1-Cyclooctylmethyl-pseudo-UTP; 1-Cyclooctyl-pseudo-UTP; 1-Cyclopentylmethyl-pseudo-UTP; 1-Cyclopentyl-pseudo-UTP; 1-Cyclopropylmethyl-pseudo-UTP; 1-Cyclopropyl-pseudo-UTP; 1-Ethyl-pseudo-UTP; 1-Hexyl-pseudo-UTP; 1-Homoallylpseudouridine TP; 1-Hydroxymethylpseudouridine TP; 1-iso-propyl-pseudo-UTP; 1-Me-2-thio-pseudo-UTP; 1-Me-4-thio-pseudo-UTP; 1-Me-alpha-thio-pseudo-UTP; 1-Methanesulfonylmethylpseudouridine TP; 1-Methoxymethylpseudouridine TP; 1-Methyl-6-(2,2,2-Trifluoroethyl)pseudo-UTP; 1-Methyl-6-(4-morpholino)-pseudo-UTP; 1-Methyl-6-(4-thiomorpholino)-pseudo-UTP; 1-Methyl-6-(substituted phenyl)pseudo-UTP; 1-Methyl-6-amino-pseudo-UTP; 1-Methyl-6-azido-pseudo-UTP; 1-Methyl-6-bromo-pseudo-UTP; 1-Methyl-6-butyl-pseudo-UTP; 1-Methyl-6-chloro-pseudo-UTP; 1-Methyl-6-cyano-pseudo-UTP; 1-Methyl-6-dimethylamino-pseudo-UTP; 1-Methyl-6-ethoxy-pseudo-UTP; 1-Methyl-6-ethylcarboxylate-pseudo-UTP; 1-Methyl-6-ethyl-pseudo-UTP; 1-Methyl-6-fluoro-pseudo-UTP; 1-Methyl-6-formyl-pseudo-UTP; 1-Methyl-6-hydroxyamino-pseudo-UTP; 1-Methyl-6-hydroxy-pseudo-UTP; 1-Methyl-6-iodo-pseudo-UTP; 1-Methyl-6-iso-propyl-pseudo-UTP; 1-Methyl-6-methoxy-pseudo-UTP; 1-Methyl-6-methylamino-pseudo-UTP; 1-Methyl-6-phenyl-pseudo-UTP; 1-Methyl-6-propyl-pseudo-UTP; 1-Methyl-6-tert-butyl-pseudo-UTP; 1-Methyl-6-trifluoromethoxy-pseudo-UTP; 1-Methyl-6-trifluoromethyl-pseudo-UTP; 1-Morpholinomethylpseudouridine TP; 1-Pentyl-pseudo-UTP; 1-Phenyl-pseudo-UTP; 1-Pivaloylpseudouridine TP; 1-Propargylpseudouridine TP; 1-Propyl-pseudo-UTP; 1-propynyl-pseudouridine; 1-p-tolyl-pseudo-UTP; 1-tert-Butyl-pseudo-UTP; 1-Thiomethoxymethylpseudouridine TP; 1-Thiomorpholinomethylpseudouridine TP; 1-Trifluoroacetylpseudouridine TP; 1-Trifluoromethyl-pseudo-UTP; 1-Vinylpseudouridine TP; 2,2′-anhydro-uridine TP; 2′-bromo-deoxyuridine TP; 2′-F-5-Methyl-2′-deoxy-UTP; 2′-OMe-5-Me-UTP; 2′-OMe-pseudo-UTP; 2′-a-Ethynyluridine TP; 2′-a-Trifluoromethyluridine TP; 2′-b-Ethynyluridine TP; 2′-b-Trifluoromethyluridine TP; 2′-Deoxy-2′,2′-difluorouridine TP; 2′-Deoxy-2′-a-mercaptouridine TP; 2′-Deoxy-2′-a-thiomethoxyuridine TP; 2′-Deoxy-2′-b-aminouridine TP; 2′-Deoxy-2′-b-azidouridine TP; 2′-Deoxy-2′-b-bromouridine TP; 2′-Deoxy-2′-b-chlorouridine TP; 2′-Deoxy-2′-b-fluorouridine TP; 2′-Deoxy-2′-b-iodouridine TP; 2′-Deoxy-2′-b-mercaptouridine TP; 2′-Deoxy-2′-b-thiomethoxyuridine TP; 2-methoxy-4-thio-uridine; 2-methoxyuridine; 2′-O-Methyl-5-(1-propynyl)uridine TP; 3-Alkyl-pseudo-UTP; 4′-Azidouridine TP; 4′-Carbocyclic uridine TP; 4′-Ethynyluridine TP; 5-(1-Propynyl)ara-uridine TP; 5-(2-Furanyl)uridine TP; 5-Cyanouridine TP; 5-Dimethylaminouridine TP; 5′-Homo-uridine TP; 5-iodo-2′-fluoro-deoxyuridine TP; 5-Phenylethynyluridine TP; 5-Trideuteromethyl-6-deuterouridine TP; 5-Trifluoromethyl-Uridine TP; 5-Vinylarauridine TP; 6-(2,2,2-Trifluoroethyl)-pseudo-UTP; 6-(4-Morpholino)-pseudo-UTP; 6-(4-Thiomorpholino)-pseudo-UTP; 6-(Substituted-Phenyl)-pseudo-UTP; 6-Amino-pseudo-UTP; 6-Azido-pseudo-UTP; 6-Bromo-pseudo-UTP; 6-Butyl-pseudo-UTP; 6-Chloro-pseudo-UTP; 6-Cyano-pseudo-UTP; 6-Dimethylamino-pseudo-UTP; 6-Ethoxy-pseudo-UTP; 6-Ethylcarboxylate-pseudo-UTP; 6-Ethyl-pseudo-UTP; 6-Fluoro-pseudo-UTP; 6-Formyl-pseudo-UTP; 6-Hydroxyamino-pseudo-UTP; 6-Hydroxy-pseudo-UTP; 6-Iodo-pseudo-UTP; 6-iso-Propyl-pseudo-UTP; 6-Methoxy-pseudo-UTP; 6-Methylamino-pseudo-UTP; 6-Methyl-pseudo-UTP; 6-Phenyl-pseudo-UTP; 6-Phenyl-pseudo-UTP; 6-Propyl-pseudo-UTP; 6-tert-Butyl-pseudo-UTP; 6-Trifluoromethoxy-pseudo-UTP; 6-Trifluoromethyl-pseudo-UTP; Alpha-thio-pseudo-UTP; Pseudouridine 1-(4-methylbenzenesulfonic acid) TP; Pseudouridine 1-(4-methylbenzoic acid) TP; Pseudouridine TP 1-[3-(2-ethoxy)]propionic acid; Pseudouridine TP 1-[3-{2-(2-[2-(2-ethoxy)-ethoxy]-ethoxy)-ethoxy}]propionic acid; Pseudouridine TP 1-[3-{2-(2-[2-{2(2-ethoxy)-ethoxy}-ethoxy]-ethoxy)-ethoxy}]propionic acid; Pseudouridine TP 1-[3-{2-(2-[2-ethoxy]-ethoxy)-ethoxy}]propionic acid; Pseudouridine TP 1-[3-{2-(2-ethoxy)-ethoxy}] propionic acid; Pseudouridine TP 1-methylphosphonic acid; Pseudouridine TP 1-methylphosphonic acid diethyl ester; Pseudo-UTP-N1-3-propionic acid; Pseudo-UTP-N1-4-butanoic acid; Pseudo-UTP-N1-5-pentanoic acid; Pseudo-UTP-N1-6-hexanoic acid; Pseudo-UTP-N1-7-heptanoic acid; Pseudo-UTP-N1-methyl-p-benzoic acid; Pseudo-UTP-N1-p-benzoic acid; Wybutosine; Hydroxywybutosine; Isowyosine; Peroxywybutosine; undermodified hydroxywybutosine; 4-demethylwyosine; 2,6-(diamino)purine; 1-(aza)-2-(thio)-3-(aza)-phenoxazin-1-yl: 1,3-(diaza)-2-(oxo)-phenthiazin-1-yl; 1,3-(diaza)-2-(oxo)-phenoxazin-1-yl; 1,3,5-(triaza)-2,6-(dioxa)-naphthalene; 2 (amino)purine; 2,4,5-(trimethyl)phenyl; 2′ methyl, 2′amino, 2′azido, 2′fluro-cytidine; 2′ methyl, 2′ amino, 2′ azido, 2′fluro-adenine; 2′methyl, 2′ amino, 2′ azido, 2′fluro-uridine; 2′-amino-2′-deoxyribose; 2-amino-6-Chloro-purine; 2-aza-inosinyl; 2′-azido-2′-deoxyribose; 2′fluoro-2′-deoxyribose; 2′-fluoro-modified bases; 2′-O-methyl-ribose; 2-oxo-7-aminopyridopyrimidin-3-yl; 2-oxo-pyridopyrimidine-3-yl; 2-pyridinone; 3 nitropyrrole; 3-(methyl)-7-(propynyl)isocarbostyrilyl; 3-(methyl)isocarbostyrilyl; 4-(fluoro)-6-(methyl)benzimidazole; 4-(methyl)benzimidazole; 4-(methyl)indolyl; 4,6-(dimethyl)indolyl; 5 nitroindole; 5 substituted pyrimidines; 5-(methyl)isocarbostyrilyl; 5-nitroindole; 6-(aza)pyrimidine; 6-(azo)thymine; 6-(methyl)-7-(aza)indolyl; 6-chloro-purine; 6-phenyl-pyrrolo-pyrimidin-2-on-3-yl; 7-(aminoalkylhydroxy)-1-(aza)-2-(thio)-3-(aza)-phenthiazin-1-yl; 7-(aminoalkylhydroxy)-1-(aza)-2-(thio)-3-(aza)-phenoxazin-1-yl; 7-(aminoalkylhydroxy)-1,3-(diaza)-2-(oxo)-phenoxazin-1-yl; 7-(aminoalkylhydroxy)-1,3-(diaza)-2-(oxo)-phenthiazin-1-yl; 7-(aminoalkylhydroxy)-1,3-(diaza)-2-(oxo)-phenoxazin-1-yl; 7-(aza)indolyl; 7-(guanidiniumalkylhydroxy)-1-(aza)-2-(thio)-3-(aza)-phenoxazinl-yl; 7-(guanidiniumalkylhydroxy)-1-(aza)-2-(thio)-3-(aza)-phenthiazin-1-yl; 7-(guanidiniumalkylhydroxy)-1-(aza)-2-(thio)-3-(aza)-phenoxazin-1-yl; 7-(guanidiniumalkylhydroxy)-1,3-(diaza)-2-(oxo)-phenoxazin-1-yl; 7-(guanidiniumalkylhydroxy)-1,3-(diaza)-2-(oxo)-phenthiazin-1-yl; 7-(guanidiniumalkylhydroxy)-1,3-(diaza)-2-(oxo)-phenoxazin-1-yl; 7-(propynyl)isocarbostyrilyl; 7-(propynyl)isocarbostyrilyl, propynyl-7-(aza)indolyl; 7-deaza-inosinyl; 7-substituted 1-(aza)-2-(thio)-3-(aza)-phenoxazin-1-yl; 7-substituted 1,3-(diaza)-2-(oxo)-phenoxazin-1-yl; 9-(methyl)-imidizopyridinyl; Aminoindolyl; Anthracenyl; bis-ortho-(aminoalkylhydroxy)-6-phenyl-pyrrolo-pyrimidin-2-on-3-yl; bis-ortho-substituted-6-phenyl-pyrrolo-pyrimidin-2-on-3-yl; Difluorotolyl; Hypoxanthine; Imidizopyridinyl; Inosinyl; Isocarbostyrilyl; Isoguanisine; N2-substituted purines; N6-methyl-2-amino-purine; N6-substituted purines; N-alkylated derivative; Napthalenyl; Nitrobenzimidazolyl; Nitroimidazolyl; Nitroindazolyl; Nitropyrazolyl; Nubularine; 06-substituted purines; O-alkylated derivative; ortho-(aminoalkylhydroxy)-6-phenyl-pyrrolo-pyrimidin-2-on-3-yl; ortho-substituted-6-phenyl-pyrrolo-pyrimidin-2-on-3-yl; Oxoformycin TP; para-(aminoalkylhydroxy)-6-phenyl-pyrrolo-pyrimidin-2-on-3-yl; para-substituted-6-phenyl-pyrrolo-pyrimidin-2-on-3-yl; Pentacenyl; Phenanthracenyl; Phenyl; propynyl-7-(aza)indolyl; Pyrenyl; pyridopyrimidin-3-yl; pyridopyrimidin-3-yl, 2-oxo-7-aminopyridopyrimidin-3-yl; pyrrolo-pyrimidin-2-on-3-yl; Pyrrolopyrimidinyl; Pyrrolopyrizinyl; Stilbenzyl; substituted 1,2,4-triazoles; Tetracenyl; Tubercidine; Xanthine; Xanthosine-5′-TP; 2-thio-zebularine; 5-aza-2-thio-zebularine; 7-deaza-2-amino-purine; pyridin-4-one ribonucleoside; 2-Amino-riboside-TP; Formycin A TP; Formycin B TP; Pyrrolosine TP; 2′-OH-ara-adenosine TP; 2′-OH-ara-cytidine TP; 2′-OH-ara-uridine TP; 2′-OH-ara-guanosine TP; 5-(2-carbomethoxyvinyl)uridine TP; and N6-(19-Amino-pentaoxanonadecyl)adenosine TP.

In some embodiments, polynucleotides (e.g., RNA polynucleotides, such as mRNA polynucleotides) include a combination of at least two (e.g., 2, 3, 4 or more) of the aforementioned modified nucleobases.

In some embodiments, modified nucleobases in polynucleotides (e.g., RNA polynucleotides, such as mRNA polynucleotides) are selected from the group consisting of pseudouridine (ψ), N1-methylpseudouridine (m¹ψ), N1-ethylpseudouridine, 2-thiouridine, 4′-thiouridine, 5-methylcytosine, 2-thio-1-methyl-1-deaza-pseudouridine, 2-thio-1-methyl-pseudouridine, 2-thio-5-aza-uridine, 2-thio-dihydropseudouridine, 2-thio-dihydrouridine, 2-thio-pseudouridine, 4-methoxy-2-thio-pseudouridine, 4-methoxy-pseudouridine, 4-thio-1-methyl-pseudouridine, 4-thio-pseudouridine, 5-aza-uridine, dihydropseudouridine, 5-methoxyuridine and 2′-O-methyl uridine. In some embodiments, polynucleotides (e.g., RNA polynucleotides, such as mRNA polynucleotides) include a combination of at least two (e.g., 2, 3, 4 or more) of the aforementioned modified nucleobases.

In some embodiments, modified nucleobases in polynucleotides (e.g., RNA polynucleotides, such as mRNA polynucleotides) are selected from the group consisting of 1-methyl-pseudouridine (m¹ψ), 5-methoxy-uridine (mo⁵U), 5-methyl-cytidine (m⁵C), pseudouridine (ψ), α-thio-guanosine and α-thio-adenosine. In some embodiments, polynucleotides (e.g., RNA polynucleotides, such as mRNA polynucleotides) include a combination of at least two (e.g., 2, 3, 4 or more) of the aforementioned modified nucleobases.

In some embodiments, polynucleotides (e.g., RNA polynucleotides, such as mRNA polynucleotides) comprise pseudouridine (ψ) and 5-methyl-cytidine (m⁵C). In some embodiments, polynucleotides (e.g., RNA polynucleotides, such as mRNA polynucleotides) comprise 1-methyl-pseudouridine (m¹ψ). In some embodiments, polynucleotides (e.g., RNA polynucleotides, such as mRNA polynucleotides) comprise 1-methyl-pseudouridine (m¹ψ) and 5-methyl-cytidine (m⁵C). In some embodiments, polynucleotides (e.g., RNA polynucleotides, such as mRNA polynucleotides) comprise 2-thiouridine (s²U). In some embodiments, polynucleotides (e.g., RNA polynucleotides, such as mRNA polynucleotides) comprise 2-thiouridine and 5-methyl-cytidine (m⁵C). In some embodiments, polynucleotides (e.g., RNA polynucleotides, such as mRNA polynucleotides) comprise methoxy-uridine (mo⁵U). In some embodiments, polynucleotides (e.g., RNA polynucleotides, such as mRNA polynucleotides) comprise 5-methoxy-uridine (mo⁵U) and 5-methyl-cytidine (m⁵C). In some embodiments, polynucleotides (e.g., RNA polynucleotides, such as mRNA polynucleotides) comprise 2′-O-methyl uridine. In some embodiments polynucleotides (e.g., RNA polynucleotides, such as mRNA polynucleotides) comprise 2′-O-methyl uridine and 5-methyl-cytidine (m⁵C). In some embodiments, polynucleotides (e.g., RNA polynucleotides, such as mRNA polynucleotides) comprise N6-methyl-adenosine (m⁶A). In some embodiments, polynucleotides (e.g., RNA polynucleotides, such as mRNA polynucleotides) comprise N6-methyl-adenosine (m⁶A) and 5-methyl-cytidine (m⁵C).

In some embodiments, polynucleotides (e.g., RNA polynucleotides, such as mRNA polynucleotides) are uniformly modified (e.g., fully modified, modified throughout the entire sequence) for a particular modification. For example, a polynucleotide can be uniformly modified with 5-methyl-cytidine (m⁵C), meaning that all cytosine residues in the mRNA sequence are replaced with 5-methyl-cytidine (m⁵C). Similarly, a polynucleotide can be uniformly modified for any type of nucleoside residue present in the sequence by replacement with a modified residue such as those set forth above.

Exemplary nucleobases and nucleosides having a modified cytosine include N4-acetyl-cytidine (ac4C), 5-methyl-cytidine (m5C), 5-halo-cytidine (e.g., 5-iodo-cytidine), 5-hydroxymethyl-cytidine (hm5C), 1-methyl-pseudoisocytidine, 2-thio-cytidine (s2C), and 2-thio-5-methyl-cytidine.

In some embodiments, a modified nucleobase is a modified uridine. In some embodiments, a modified nucleobase is a modified cytosine. Nucleosides having a modified uridine include 5-cyano uridine, and 4′-thio uridine.

In some embodiments, a modified nucleobase is a modified adenine. Exemplary nucleobases and nucleosides having a modified adenine include 7-deaza-adenine, 1-methyl-adenosine (m1A), 2-methyl-adenine (m2A), and N6-methyl-adenosine (m6A).

In some embodiments, a modified nucleobase is a modified guanine. Exemplary nucleobases and nucleosides having a modified guanine include inosine (I), 1-methyl-inosine (m1I), wyosine (imG), methylwyosine (mimG), 7-deaza-guanosine, 7-cyano-7-deaza-guanosine (preQ0), 7-aminomethyl-7-deaza-guanosine (preQ1), 7-methyl-guanosine (m7G), 1-methyl-guanosine (m1G), 8-oxo-guanosine, 7-methyl-8-oxo-guanosine.

The polynucleotides of the present disclosure may be partially or fully modified along the entire length of the molecule. For example, one or more or all or a given type of nucleotide (e.g., purine or pyrimidine, or any one or more or all of A, G, U, C) may be uniformly modified in a polynucleotide of the invention, or in a given predetermined sequence region thereof (e.g., in the mRNA including or excluding the polyA tail). In some embodiments, all nucleotides X in a polynucleotide of the present disclosure (or in a given sequence region thereof) are modified nucleotides, wherein X may any one of nucleotides A, G, U, C, or any one of the combinations A+G, A+U, A+C, G+U, G+C, U+C, A+G+U, A+G+C, G+U+C or A+G+C.

The polynucleotide may contain from about 1% to about 100% modified nucleotides (either in relation to overall nucleotide content, or in relation to one or more types of nucleotide, i.e., any one or more of A, G, U or C) or any intervening percentage (e.g., from 1% to 20%, from 1% to 25%, from 1% to 50%, from 1% to 60%, from 1% to 70%, from 1% to 80%, from 1% to 90%, from 1% to 95%, from 10% to 20%, from 10% to 25%, from 10% to 50%, from 10% to 60%, from 10% to 70%, from 10% to 80%, from 10% to 90%, from 10% to 95%, from 10% to 100%, from 20% to 25%, from 20% to 50%, from 20% to 60%, from 20% to 70%, from 20% to 80%, from 20% to 90%, from 20% to 95%, from 20% to 100%, from 50% to 60%, from 50% to 70%, from 50% to 80%, from 50% to 90%, from 50% to 95%, from 50% to 100%, from 70% to 80%, from 70% to 90%, from 70% to 95%, from 70% to 100%, from 80% to 90%, from 80% to 95%, from 80% to 100%, from 90% to 95%, from 90% to 100%, and from 95% to 100%). Any remaining percentage is accounted for by the presence of unmodified A, G, U, or C.

The polynucleotides may contain at a minimum 1% and at maximum 100% modified nucleotides, or any intervening percentage, such as at least 5% modified nucleotides, at least 10% modified nucleotides, at least 25% modified nucleotides, at least 50% modified nucleotides, at least 80% modified nucleotides, or at least 90% modified nucleotides. For example, the polynucleotides may contain a modified pyrimidine such as a modified uracil or cytosine. In some embodiments, at least 5%, at least 10%, at least 25%, at least 50%, at least 80%, at least 90% or 100% of the uracil in the polynucleotide is replaced with a modified uracil (e.g., a 5-substituted uracil). The modified uracil can be replaced by a compound having a single unique structure, or can be replaced by a plurality of compounds having different structures (e.g., 2, 3, 4 or more unique structures). In some embodiments, at least 5%, at least 10%, at least 25%, at least 50%, at least 80%, at least 90% or 100% of the cytosine in the polynucleotide is replaced with a modified cytosine (e.g., a 5-substituted cytosine). The modified cytosine can be replaced by a compound having a single unique structure, or can be replaced by a plurality of compounds having different structures (e.g., 2, 3, 4 or more unique structures).

Thus, in some embodiments, the RNA (e.g., mRNA) vaccines comprise a 5′UTR element, an optionally codon optimized open reading frame, and a 3′UTR element, a poly(A) sequence and/or a polyadenylation signal wherein the RNA is not chemically modified.

In some embodiments, the modified nucleobase is a modified uracil. Exemplary nucleobases and nucleosides having a modified uracil include pseudouridine (ψ), pyridin-4-one ribonucleoside, 5-aza-uridine, 6-aza-uridine, 2-thio-5-aza-uridine, 2-thio-uridine (s²U), 4-thio-uridine (s⁴U), 4-thio-pseudouridine, 2-thio-pseudouridine, 5-hydroxy-uridine (ho⁵U), 5-aminoallyl-uridine, 5-halo-uridine (e.g., 5-iodo-uridineor 5-bromo-uridine), 3-methyl-uridine (m³U), 5-methoxy-uridine (mo⁵U), uridine 5-oxyacetic acid (cmo⁵U), uridine 5-oxyacetic acid methyl ester (mcmo⁵U), 5-carboxymethyl-uridine (cm⁵U), 1-carboxymethyl-pseudouridine, 5-carboxyhydroxymethyl-uridine (chm⁵U), 5-carboxyhydroxymethyl-uridine methyl ester (mchm⁵U), 5-methoxycarbonylmethyl-uridine (mcm⁵U), 5-methoxycarbonylmethyl-2-thio-uridine (mcm⁵s²U), 5-aminomethyl-2-thio-uridine (nm⁵s²U), 5-methylaminomethyl-uridine (mnm⁵U), 5-methylaminomethyl-2-thio-uridine (mnm⁵s²U), 5-methylaminomethyl-2-seleno-uridine (mnm⁵se²U), 5-carbamoylmethyl-uridine (ncm⁵U), 5-carboxymethylaminomethyl-uridine (cmnm⁵U), 5-carboxymethylaminomethyl-2-thio-uridine (cmnm⁵s²U), 5-propynyl-uridine, 1-propynyl-pseudouridine, 5-taurinomethyl-uridine (τm⁵U), 1-taurinomethyl-pseudouridine, 5-taurinomethyl-2-thio-uridine(τm⁵s²U), 1-taurinomethyl-4-thio-pseudouridine, 5-methyl-uridine (m⁵U, i.e., having the nucleobase deoxythymine), 1-methyl-pseudouridine (m¹ψ), 5-methyl-2-thio-uridine (m⁵s²U), 1-methyl-4-thio-pseudouridine (m¹s⁴ψ), 4-thio-1-methyl-pseudouridine, 3-methyl-pseudouridine (m³ψ), 2-thio-1-methyl-pseudouridine, 1-methyl-1-deaza-pseudouridine, 2-thio-1-methyl-1-deaza-pseudouridine, dihydrouridine (D), dihydropseudouridine, 5,6-dihydrouridine, 5-methyldihydrouridine (m⁵D), 2-thio-dihydrouridine, 2-thio-dihydropseudouridine, 2-methoxy-uridine, 2-methoxy-4-thio-uridine, 4-methoxy-pseudouridine, 4-methoxy-2-thio-pseudouridine, N1-methyl-pseudouridine, 3-(3-amino-3-carboxypropyl)uridine (acp³U), 1-methyl-3-(3-amino-3-carboxypropyl)pseudouridine (acp³ψ), 5-(isopentenylaminomethyl)uridine (inm⁵U), 5-(isopentenylaminomethyl)-2-thio-uridine (inm⁵s²U), α-thio-uridine, 2′-O-methyl-uridine (Um), 5,2′-O-dimethyl-uridine (m⁵Um), 2′-O-methyl-pseudouridine (ψm), 2-thio-2′-O-methyl-uridine (s²Um), 5-methoxycarbonylmethyl-2′-O-methyl-uridine (mcm⁵Um), 5-carbamoylmethyl-2′-O-methyl-uridine (ncm⁵Um), 5-carboxymethylaminomethyl-2′-O-methyl-uridine (cmnm⁵Um), 3,2′-O-dimethyl-uridine (m³Um), and 5-(isopentenylaminomethyl)-2′-O-methyl-uridine (inm⁵Um), 1-thio-uridine, deoxythymidine, 2′-F-ara-uridine, 2′-F-uridine, 2′-OH-ara-uridine, 5-(2-carbomethoxyvinyl) uridine, and 5-[3-(1-E-propenylamino)]uridine.

In some embodiments, the modified nucleobase is a modified cytosine. Exemplary nucleobases and nucleosides having a modified cytosine include 5-aza-cytidine, 6-aza-cytidine, pseudoisocytidine, 3-methyl-cytidine (m³C), N4-acetyl-cytidine (ac⁴C), 5-formylcytidine (f⁵C), N4-methyl-cytidine (m⁴C), 5-methyl-cytidine (m⁵C), 5-halo-cytidine (e.g., 5-iodo-cytidine), 5-hydroxymethyl-cytidine (hm⁵C), 1-methyl-pseudoisocytidine, pyrrolo-cytidine, pyrrolo-pseudoisocytidine, 2-thio-cytidine (s²C), 2-thio-5-methyl-cytidine, 4-thio-pseudoisocytidine, 4-thio-1-methyl-pseudoisocytidine, 4-thio-1-methyl-1-deaza-pseudoisocytidine, 1-methyl-1-deaza-pseudoisocytidine, zebularine, 5-aza-zebularine, 5-methyl-zebularine, 5-aza-2-thio-zebularine, 2-thio-zebularine, 2-methoxy-cytidine, 2-methoxy-5-methyl-cytidine, 4-methoxy-pseudoisocytidine, 4-methoxy-1-methyl-pseudoisocytidine, lysidine (k₂C), α-thio-cytidine, 2′-O-methyl-cytidine (Cm), 5,2′-O-dimethyl-cytidine (m⁵Cm), N4-acetyl-2′-O-methyl-cytidine (ac⁴Cm), N4,2′-O-dimethyl-cytidine (m⁴Cm), 5-formyl-2′-O-methyl-cytidine (f⁵Cm), N4,N4,2′-O-trimethyl-cytidine (m⁴ ₂ Cm), 1-thio-cytidine, 2′-F-ara-cytidine, 2′-F-cytidine, and 2′-OH-ara-cytidine.

In some embodiments, the modified nucleobase is a modified adenine. Exemplary nucleobases and nucleosides having a modified adenine include 2-amino-purine, 2, 6-diaminopurine, 2-amino-6-halo-purine (e.g., 2-amino-6-chloro-purine), 6-halo-purine (e.g., 6-chloro-purine), 2-amino-6-methyl-purine, 8-azido-adenosine, 7-deaza-adenine, 7-deaza-8-aza-adenine, 7-deaza-2-amino-purine, 7-deaza-8-aza-2-amino-purine, 7-deaza-2,6-diaminopurine, 7-deaza-8-aza-2,6-diaminopurine, 1-methyl-adenosine (m¹A), 2-methyl-adenine (m²A), N6-methyl-adenosine (m⁶A), 2-methylthio-N6-methyl-adenosine (ms² m⁶A), N6-isopentenyl-adenosine (i⁶A), 2-methylthio-N6-isopentenyl-adenosine (ms²i⁶A), N6-(cis-hydroxyisopentenyl)adenosine (io⁶A), 2-methylthio-N6-(cis-hydroxyisopentenyl)adenosine (ms²io⁶A), N6-glycinylcarbamoyl-adenosine (g⁶A), N6-threonylcarbamoyl-adenosine (t⁶A), N6-methyl-N6-threonylcarbamoyl-adenosine (m⁶t⁶A), 2-methylthio-N6-threonylcarbamoyl-adenosine (ms²g⁶A), N6,N6-dimethyl-adenosine (m⁶ ₂A), N6-hydroxynorvalylcarbamoyl-adenosine (hn⁶A), 2-methylthio-N6-hydroxynorvalylcarbamoyl-adenosine (ms²hn⁶A), N6-acetyl-adenosine (ac⁶A), 7-methyl-adenine, 2-methylthio-adenine, 2-methoxy-adenine, α-thio-adenosine, 2′-O-methyl-adenosine (Am), N6,2′-O-dimethyl-adenosine (m⁶Am), N6,N6,2′-O-trimethyl-adenosine (m⁶ ₂ Am), 1,2′-O-dimethyl-adenosine (m¹Am), 2′-O-ribosyladenosine (phosphate) (Ar(p)), 2-amino-N6-methyl-purine, 1-thio-adenosine, 8-azido-adenosine, 2′-F-ara-adenosine, 2′-F-adenosine, 2′-OH-ara-adenosine, and N6-(19-amino-pentaoxanonadecyl)-adenosine.

In some embodiments, the modified nucleobase is a modified guanine. Exemplary nucleobases and nucleosides having a modified guanine include inosine (I), 1-methyl-inosine (m¹I), wyosine (imG), methylwyosine (mimG), 4-demethyl-wyosine (imG-14), isowyosine (imG2), wybutosine (yW), peroxywybutosine (o₂yW), hydroxywybutosine (OhyW), undermodified hydroxywybutosine (OhyW*), 7-deaza-guanosine, queuosine (Q), epoxyqueuosine (oQ), galactosyl-queuosine (galQ), mannosyl-queuosine (manQ), 7-cyano-7-deaza-guanosine (preQ₀), 7-aminomethyl-7-deaza-guanosine (preQ₁), archaeosine (G⁺), 7-deaza-8-aza-guanosine, 6-thio-guanosine, 6-thio-7-deaza-guanosine, 6-thio-7-deaza-8-aza-guanosine, 7-methyl-guanosine (m⁷G), 6-thio-7-methyl-guanosine, 7-methyl-inosine, 6-methoxy-guanosine, 1-methyl-guanosine (m¹G), N2-methyl-guanosine (m²G), N2,N2-dimethyl-guanosine (m² ₂ G), N2,7-dimethyl-guano sine (m^(2,7)G), N2, N2,7-dimethyl-guanosine (m^(2,2,7)G), 8-oxo-guanosine, 7-methyl-8-oxo-guanosine, 1-methyl-6-thio-guanosine, N2-methyl-6-thio-guanosine, N2,N2-dimethyl-6-thio-guanosine, α-thio-guanosine, 2′-O-methyl-guanosine (Gm), N2-methyl-2′-O-methyl-guanosine (m²Gm), N2,N2-dimethyl-2′-O-methyl-guanosine (m² ₂ Gm), 1-methyl-2′-O-methyl-guanosine (m¹Gm), N2,7-dimethyl-2′-O-methyl-guanosine (m^(2,7)Gm), 2′-O-methyl-inosine (Im), 1,2′-O-dimethyl-inosine (m¹Im), 2′-O-ribosylguanosine (phosphate) (Gr(p)), 1-thio-guanosine, 06-methyl-guano sine, 2′-F-ara-guanosine, and 2′-F-guanosine.

N-Linked Glycosylation Site Mutants

N-linked glycans of viral proteins play important roles in modulating the immune response. Glycans can be important for maintaining the appropriate antigenic conformations, shielding potential neutralization epitopes, and may alter the proteolytic susceptibility of proteins. Some viruses have putative N-linked glycosylation sites. Deletion or modification of an N-linked glycosylation site may enhance the immune response. Thus, the present disclosure provides, in some embodiments, RNA (e.g., mRNA) vaccines comprising nucleic acids (e.g., mRNA) encoding antigenic polypeptides that comprise a deletion or modification at one or more N-linked glycosylation sites.

In Vitro Transcription of RNA (e.g., mRNA)

Tropical disease vaccines of the present disclosure comprise at least one RNA polynucleotide, such as a mRNA (e.g., modified mRNA). mRNA, for example, is transcribed in vitro from template DNA, referred to as an “in vitro transcription template.” In some embodiments, an in vitro transcription template encodes a 5′ untranslated (UTR) region, contains an open reading frame, and encodes a 3′ UTR and a polyA tail. The particular nucleic acid sequence composition and length of an in vitro transcription template will depend on the mRNA encoded by the template.

A “5′ untranslated region” (5′UTR) refers to a region of an mRNA that is directly upstream (i.e., 5′) from the start codon (i.e., the first codon of an mRNA transcript translated by a ribosome) that does not encode a polypeptide.

A “3′ untranslated region” (3′UTR) refers to a region of an mRNA that is directly downstream (i.e., 3′) from the stop codon (i.e., the codon of an mRNA transcript that signals a termination of translation) that does not encode a polypeptide.

An “open reading frame” is a continuous stretch of DNA beginning with a start codon (e.g., methionine (ATG)), and ending with a stop codon (e.g., TAA, TAG or TGA) and encodes a polypeptide.

A “polyA tail” is a region of mRNA that is downstream, e.g., directly downstream (i.e., 3′), from the 3′ UTR that contains multiple, consecutive adenosine monophosphates. A polyA tail may contain 10 to 300 adenosine monophosphates. For example, a polyA tail may contain 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290 or 300 adenosine monophosphates. In some embodiments, a polyA tail contains 50 to 250 adenosine monophosphates. In a relevant biological setting (e.g., in cells, in vivo) the poly(A) tail functions to protect mRNA from enzymatic degradation, e.g., in the cytoplasm, and aids in transcription termination, export of the mRNA from the nucleus and translation.

In some embodiments, a polynucleotide includes 200 to 3,000 nucleotides. For example, a polynucleotide may include 200 to 500, 200 to 1000, 200 to 1500, 200 to 3000, 500 to 1000, 500 to 1500, 500 to 2000, 500 to 3000, 1000 to 1500, 1000 to 2000, 1000 to 3000, 1500 to 3000, or 2000 to 3000 nucleotides.

Flagellin Adjuvants

Flagellin is an approximately 500 amino acid monomeric protein that polymerizes to form the flagella associated with bacterial motion. Flagellin is expressed by a variety of flagellated bacteria (Salmonella typhimurium for example) as well as non-flagellated bacteria (such as Escherichia coli). Sensing of flagellin by cells of the innate immune system (dendritic cells, macrophages, etc.) is mediated by the Toll-like receptor 5 (TLR5) as well as by Nod-like receptors (NLRs) Ipaf and Naip5. TLRs and NLRs have been identified as playing a role in the activation of innate immune response and adaptive immune response. As such, flagellin provides an adjuvant effect in a vaccine.

The nucleotide and amino acid sequences encoding known flagellin polypeptides are publicly available in the NCBI GenBank database. The flagellin sequences from S. Typhimurium, H. Pylori, V. Cholera, S. marcesens, S. flexneri, T. pallidum, L. pneumophila, B. burgdorferei, C. difficile, R. meliloti, A. tumefaciens, R. lupini, B. clarridgeiae, P. Mirabilis, B. subtilus, L. monocytogenes, P. aeruginosa, and E. coli, among others are known.

A flagellin polypeptide, as used herein, refers to a full length flagellin protein, immunogenic fragments thereof, and peptides having at least 50% sequence identity to a flagellin protein or immunogenic fragments thereof. Exemplary flagellin proteins include flagellin from Salmonella typhi (UniPro Entry number: Q56086), Salmonella typhimurium (A0A0C9DG09), Salmonella enteritidis (A0A0C9BAB7), and Salmonella choleraesuis (Q6V2X8), and proteins having an amino acid sequence identified by any one of SEQ ID NO: 420-422 (Table 66). In some embodiments, the flagellin polypeptide has at least 60%, 70%, 75%, 80%, 90%, 95%, 97%, 98%, or 99% sequence identity to a flagellin protein or immunogenic fragments thereof.

In some embodiments, the flagellin polypeptide is an immunogenic fragment. An immunogenic fragment is a portion of a flagellin protein that provokes an immune response. In some embodiments, the immune response is a TLR5 immune response. An example of an immunogenic fragment is a flagellin protein in which all or a portion of a hinge region has been deleted or replaced with other amino acids. For example, an antigenic polypeptide may be inserted in the hinge region. Hinge regions are the hypervariable regions of a flagellin. Hinge regions of a flagellin are also referred to as “D3 domain or region,” “propeller domain or region,” “hypervariable domain or region” and “variable domain or region.” “At least a portion of a hinge region,” as used herein, refers to any part of the hinge region of the flagellin, or the entirety of the hinge region. In other embodiments an immunogenic fragment of flagellin is a 20, 25, 30, 35, or 40 amino acid C-terminal fragment of flagellin.

The flagellin monomer is formed by domains D0 through D3. D0 and D1, which form the stem, are composed of tandem long alpha helices and are highly conserved among different bacteria. The D1 domain includes several stretches of amino acids that are useful for TLR5 activation. The entire D1 domain or one or more of the active regions within the domain are immunogenic fragments of flagellin. Examples of immunogenic regions within the D1 domain include residues 88-114 and residues 411-431 in Salmonella typhimurium FliC flagellin. Within the 13 amino acids in the 88-100 region, at least 6 substitutions are permitted between Salmonella flagellin and other flagellins that still preserve TLR5 activation. Thus, immunogenic fragments of flagellin include flagellin like sequences that activate TLR5 and contain a 13 amino acid motif that is 53% or more identical to the Salmonella sequence in 88-100 of FliC (LQRVRELAVQSAN; SEQ ID NO: 428).

In some embodiments, the RNA (e.g., mRNA) vaccine includes an RNA that encodes a fusion protein of flagellin and one or more antigenic polypeptides. A “fusion protein” as used herein, refers to a linking of two components of the construct. In some embodiments, a carboxy-terminus of the antigenic polypeptide is fused or linked to an amino terminus of the flagellin polypeptide. In other embodiments, an amino-terminus of the antigenic polypeptide is fused or linked to a carboxy-terminus of the flagellin polypeptide. The fusion protein may include, for example, one, two, three, four, five, six or more flagellin polypeptides linked to one, two, three, four, five, six or more antigenic polypeptides. When two or more flagellin polypeptides and/or two or more antigenic polypeptides are linked such a construct may be referred to as a “multimer.”

Each of the components of a fusion protein may be directly linked to one another or they may be connected through a linker. For instance, the linker may be an amino acid linker. The amino acid linker encoded for by the RNA (e.g., mRNA) vaccine to link the components of the fusion protein may include, for instance, at least one member selected from the group consisting of a lysine residue, a glutamic acid residue, a serine residue and an arginine residue. In some embodiments the linker is 1-30, 1-25, 1-25, 5-10, 5, 15, or 5-20 amino acids in length.

In other embodiments the RNA (e.g., mRNA) vaccine includes at least two separate RNA polynucleotides, one encoding one or more antigenic polypeptides and the other encoding the flagellin polypeptide. The at least two RNA polynucleotides may be coformulated in a carrier such as a lipid nanoparticle.

Broad Spectrum RNA (e.g., mRNA) Vaccines

There may be situations where persons are at risk for infection with more than one strain of Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV. RNA (e.g., mRNA) therapeutic vaccines are particularly amenable to combination vaccination approaches due to a number of factors including, but not limited to, speed of manufacture, ability to rapidly tailor vaccines to accommodate perceived geographical threat, and the like. Moreover, because the vaccines utilize the human body to produce the antigenic protein, the vaccines are amenable to the production of larger, more complex antigenic proteins, allowing for proper folding, surface expression, antigen presentation, etc. in the human subject. To protect against more than one strain of Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV, a combination vaccine can be administered that includes RNA (e.g., mRNA) encoding at least one antigenic polypeptide protein (or antigenic portion thereof) of a first tropical disease virus or organism and further includes RNA encoding at least one antigenic polypeptide protein (or antigenic portion thereof) of a second tropical disease virus or organism. RNA (e.g., mRNA) can be coformulated, for example, in a single lipid nanoparticle (LNP) or can be formulated in separate LNPs for co-administration.

Methods of Treatment

Provided herein are compositions (e.g., pharmaceutical compositions), methods, kits and reagents for prevention and/or treatment of tropical diseases in humans and other mammals. Tropical disease RNA (e.g. mRNA) vaccines can be used as therapeutic or prophylactic agents, alone or in combination with other vaccine(s). They may be used in medicine to prevent and/or treat tropical disease. In exemplary aspects, the RNA (e.g., mRNA) vaccines of the present disclosure are used to provide prophylactic protection from Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV. Prophylactic protection from Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV can be achieved following administration of a RNA (e.g., mRNA) vaccine of the present disclosure. Tropical disease RNA (e.g., mRNA) vaccines of the present disclosure may be used to treat or prevent viral “co-infections” containing two or more tropical disease infections. Vaccines can be administered once, twice, three times, four times or more, but it is likely sufficient to administer the vaccine once (optionally followed by a single booster). It is possible, although less desirable, to administer the vaccine to an infected individual to achieve a therapeutic response. Dosing may need to be adjusted accordingly.

A method of eliciting an immune response in a subject against Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV is provided in aspects of the present disclosure. The method involves administering to the subject a tropical disease RNA (e.g., mRNA) vaccine comprising at least one RNA (e.g., mRNA) polynucleotide having an open reading frame encoding at least one Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide, thereby inducing in the subject an immune response specific to Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide or an immunogenic fragment thereof, wherein anti-antigenic polypeptide antibody titer in the subject is increased following vaccination relative to anti-antigenic polypeptide antibody titer in a subject vaccinated with a prophylactically effective dose of a traditional vaccine against Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV. An “anti-antigenic polypeptide antibody” is a serum antibody the binds specifically to the antigenic polypeptide.

In some embodiments, a RNA (e.g., mRNA) vaccine (e.g., a Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV RNA vaccine) capable of eliciting an immune response is administered intramuscularly or intranasally via a composition including a compound according to Formula (I), (IA), (II), (IIa), (IIb), (IIc), (IId) or (IIe) (e.g., Compound 3, 18, 20, 25, 26, 29, 30, 60, 108-112, or 122).

A prophylactically effective dose is a therapeutically effective dose that prevents infection with the virus at a clinically acceptable level. In some embodiments the therapeutically effective dose is a dose listed in a package insert for the vaccine. A traditional vaccine, as used herein, refers to a vaccine other than the RNA (e.g., mRNA) vaccines of the present disclosure. For instance, a traditional vaccine includes but is not limited to live/attenuated microorganism vaccines, killed/inactivated microorganism vaccines, subunit vaccines, protein antigen vaccines, DNA vaccines, VLP vaccines, etc. In exemplary embodiments, a traditional vaccine is a vaccine that has achieved regulatory approval and/or is registered by a national drug regulatory body, for example the Food and Drug Administration (FDA) in the United States or the European Medicines Agency (EMA).

In some embodiments the anti-antigenic polypeptide antibody titer in the subject is increased 1 log to 10 log following vaccination relative to anti-antigenic polypeptide antibody titer in a subject vaccinated with a prophylactically effective dose of a traditional vaccine against Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV.

In some embodiments the anti-antigenic polypeptide antibody titer in the subject is increased 1 log, 2 log, 3 log, 5 log or 10 log following vaccination relative to anti-antigenic polypeptide antibody titer in a subject vaccinated with a prophylactically effective dose of a traditional vaccine against Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV.

A method of eliciting an immune response in a subject against Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV is provided in other aspects of the disclosure. The method involves administering to the subject a tropical disease RNA (e.g., mRNA) vaccine comprising at least one RNA (e.g., mRNA) polynucleotide having an open reading frame encoding at least one Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide or an immunogenic fragment thereof, thereby inducing in the subject an immune response specific to Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide or an immunogenic fragment thereof, wherein the immune response in the subject is equivalent to an immune response in a subject vaccinated with a traditional vaccine against the Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV at 2 times to 100 times the dosage level relative to the RNA (e.g., mRNA) vaccine.

In some embodiments, the immune response in the subject is equivalent to an immune response in a subject vaccinated with a traditional vaccine at 2, 3, 4, 5, 10, 50, 100 times the dosage level relative to the Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV RNA (e.g., mRNA) vaccine.

In some embodiments the immune response in the subject is equivalent to an immune response in a subject vaccinated with a traditional vaccine at 10-100 times, or 100-1000 times, the dosage level relative to the Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV RNA (e.g., mRNA) vaccine.

In some embodiments the immune response is assessed by determining protein antibody titer in the subject.

Some embodiments provide a method of inducing an immune response in a subject by administering to the subject a tropical disease RNA (e.g., mRNA) vaccine comprising at least one RNA (e.g., mRNA) polynucleotide having an open reading frame encoding at least one Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide, thereby inducing in the subject an immune response specific to the antigenic polypeptide or an immunogenic fragment thereof, wherein the immune response in the subject is induced 2 days to 10 weeks earlier relative to an immune response induced in a subject vaccinated with a prophylactically effective dose of a traditional vaccine against Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV. In some embodiments, the immune response in the subject is induced in a subject vaccinated with a prophylactically effective dose of a traditional vaccine at 2 times to 100 times the dosage level relative to the RNA (e.g., mRNA) vaccine.

In some embodiments the immune response in the subject is equivalent to an immune response in a subject vaccinated with a traditional vaccine at 2, 3, 4, 5, 10, 50, 100 times the dosage level relative to the Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV RNA (e.g., mRNA) vaccine.

In some embodiments, the immune response in the subject is induced 2 days earlier, or 3 days earlier, relative to an immune response induced in a subject vaccinated with a prophylactically effective dose of a traditional vaccine.

In some embodiments the immune response in the subject is induced 1 week, 2 weeks, 3 weeks, 5 weeks, or 10 weeks earlier relative to an immune response induced in a subject vaccinated with a prophylactically effective dose of a traditional vaccine.

Therapeutic and Prophylactic Compositions

Provided herein are compositions (e.g., pharmaceutical compositions), methods, kits and reagents for prevention, treatment or diagnosis of Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV in humans and other mammals, for example. Tropical disease RNA (e.g. mRNA) vaccines can be used as therapeutic or prophylactic agents. They may be used in medicine to prevent and/or treat infectious disease. In some embodiments, the RNA (e.g., mRNA) vaccines of the present disclosure are used fin the priming of immune effector cells, for example, to activate peripheral blood mononuclear cells (PBMCs) ex vivo, which are then infused (re-infused) into a subject.

In some embodiments, tropical disease vaccine containing RNA (e.g., mRNA) polynucleotides as described herein can be administered to a subject (e.g., a mammalian subject, such as a human subject), and the RNA (e.g., mRNA) polynucleotides are translated in vivo to produce an antigenic polypeptide.

The tropical disease RNA (e.g., mRNA) vaccines may be induced for translation of a polypeptide (e.g., antigen or immunogen) in a cell, tissue or organism. In some embodiments, such translation occurs in vivo, although such translation may occur ex vivo, in culture or in vitro. In some embodiments, the cell, tissue or organism is contacted with an effective amount of a composition containing a tropical disease RNA (e.g., mRNA) vaccine that contains a polynucleotide that has at least one a translatable region encoding an antigenic polypeptide.

An “effective amount” of a tropical disease RNA (e.g. mRNA) vaccine is provided based, at least in part, on the target tissue, target cell type, means of administration, physical characteristics of the polynucleotide (e.g., size, and extent of modified nucleosides) and other components of the vaccine, and other determinants. In general, an effective amount of the tropical disease RNA (e.g., mRNA) vaccine composition provides an induced or boosted immune response as a function of antigen production in the cell, preferably more efficient than a composition containing a corresponding unmodified polynucleotide encoding the same antigen or a peptide antigen. Increased antigen production may be demonstrated by increased cell transfection (the percentage of cells transfected with the RNA, e.g., mRNA, vaccine), increased protein translation from the polynucleotide, decreased nucleic acid degradation (as demonstrated, for example, by increased duration of protein translation from a modified polynucleotide), or altered antigen specific immune response of the host cell.

In some embodiments, RNA (e.g. mRNA) vaccines (including polynucleotides their encoded polypeptides) in accordance with the present disclosure may be used for treatment of Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV.

Tropical disease RNA (e.g. mRNA) vaccines may be administered prophylactically or therapeutically as part of an active immunization scheme to healthy individuals or early in infection during the incubation phase or during active infection after onset of symptoms. In some embodiments, the amount of RNA (e.g., mRNA) vaccine of the present disclosure provided to a cell, a tissue or a subject may be an amount effective for immune prophylaxis.

Tropical disease RNA (e.g. mRNA) vaccines may be administrated with other prophylactic or therapeutic compounds. As a non-limiting example, a prophylactic or therapeutic compound may be an adjuvant or a booster. As used herein, when referring to a prophylactic composition, such as a vaccine, the term “booster” refers to an extra administration of the prophylactic (vaccine) composition. A booster (or booster vaccine) may be given after an earlier administration of the prophylactic composition. The time of administration between the initial administration of the prophylactic composition and the booster may be, but is not limited to, 1 minute, 2 minutes, 3 minutes, 4 minutes, 5 minutes, 6 minutes, 7 minutes, 8 minutes, 9 minutes, 10 minutes, 15 minutes, 20 minutes 35 minutes, 40 minutes, 45 minutes, 50 minutes, 55 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 1 day, 36 hours, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 10 days, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 18 months, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years, 11 years, 12 years, 13 years, 14 years, 15 years, 16 years, 17 years, 18 years, 19 years, 20 years, 25 years, 30 years, 35 years, 40 years, 45 years, 50 years, 55 years, 60 years, 65 years, 70 years, 75 years, 80 years, 85 years, 90 years, 95 years or more than 99 years. In some embodiments, the time of administration between the initial administration of the prophylactic composition and the booster may be, but is not limited to, 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 6 months or 1 year.

In some embodiments, tropical disease RNA (e.g. mRNA) vaccines may be administered intramuscularly, intradermally, or intranasally, similarly to the administration of inactivated vaccines known in the art.

Tropical disease RNA (e.g. mRNA) vaccines may be utilized in various settings depending on the prevalence of the infection or the degree or level of unmet medical need. As a non-limiting example, the RNA (e.g., mRNA) vaccines may be utilized to treat and/or prevent a variety of tropical diseases. RNA (e.g., mRNA) vaccines have superior properties in that they produce much larger antibody titers and produce responses early than commercially available anti-viral agents/compositions.

Provided herein are pharmaceutical compositions including tropical disease RNA (e.g. mRNA) vaccines and RNA (e.g. mRNA) vaccine compositions and/or complexes optionally in combination with one or more pharmaceutically acceptable excipients.

Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV RNA (e.g. mRNA) vaccines may be formulated or administered alone or in conjunction with one or more other components. For instance, Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV RNA (e.g., mRNA) vaccines (vaccine compositions) may comprise other components including, but not limited to, adjuvants.

In some embodiments, tropical disease (e.g. mRNA) vaccines do not include an adjuvant (they are adjuvant free).

Tropical disease RNA (e.g. mRNA) vaccines may be formulated or administered in combination with one or more pharmaceutically-acceptable excipients. In some embodiments, vaccine compositions comprise at least one additional active substances, such as, for example, a therapeutically-active substance, a prophylactically-active substance, or a combination of both. Vaccine compositions may be sterile, pyrogen-free or both sterile and pyrogen-free. General considerations in the formulation and/or manufacture of pharmaceutical agents, such as vaccine compositions, may be found, for example, in Remington: The Science and Practice of Pharmacy 21st ed., Lippincott Williams & Wilkins, 2005 (incorporated herein by reference in its entirety).

In some embodiments, tropical disease RNA (e.g. mRNA) vaccines are administered to humans, human patients or subjects. For the purposes of the present disclosure, the phrase “active ingredient” generally refers to the RNA (e.g., mRNA) vaccines or the polynucleotides contained therein, for example, RNA polynucleotides (e.g., mRNA polynucleotides) encoding antigenic polypeptides.

Formulations of the tropical disease vaccine compositions described herein may be prepared by any method known or hereafter developed in the art of pharmacology. In general, such preparatory methods include the step of bringing the active ingredient (e.g., mRNA polynucleotide) into association with an excipient and/or one or more other accessory ingredients, and then, if necessary and/or desirable, dividing, shaping and/or packaging the product into a desired single- or multi-dose unit.

Relative amounts of the active ingredient, the pharmaceutically acceptable excipient, and/or any additional ingredients in a pharmaceutical composition in accordance with the disclosure will vary, depending upon the identity, size, and/or condition of the subject treated and further depending upon the route by which the composition is to be administered. By way of example, the composition may comprise between 0.1% and 100%, e.g., between 0.5 and 50%, between 1-30%, between 5-80%, at least 80% (w/w) active ingredient.

Tropical disease RNA (e.g. mRNA) vaccines can be formulated using one or more excipients to: increase stability; increase cell transfection; permit the sustained or delayed release (e.g., from a depot formulation); alter the biodistribution (e.g., target to specific tissues or cell types); increase the translation of encoded protein in vivo; and/or alter the release profile of encoded protein (antigen) in vivo. In addition to traditional excipients such as any and all solvents, dispersion media, diluents, or other liquid vehicles, dispersion or suspension aids, surface active agents, isotonic agents, thickening or emulsifying agents, preservatives, excipients can include, without limitation, lipidoids, liposomes, lipid nanoparticles, polymers, lipoplexes, core-shell nanoparticles, peptides, proteins, cells transfected with tropical disease RNA (e.g. mRNA) vaccines (e.g., for transplantation into a subject), hyaluronidase, nanoparticle mimics and combinations thereof.

Stabilizing Elements

Naturally-occurring eukaryotic mRNA molecules have been found to contain stabilizing elements, including, but not limited to untranslated regions (UTR) at their 5′-end (5′UTR) and/or at their 3′-end (3′UTR), in addition to other structural features, such as a 5′-cap structure or a 3′-poly(A) tail. Both the 5′UTR and the 3′UTR are typically transcribed from the genomic DNA and are elements of the premature mRNA. Characteristic structural features of mature mRNA, such as the 5′-cap and the 3′-poly(A) tail are usually added to the transcribed (premature) mRNA during mRNA processing. The 3′-poly(A) tail is typically a stretch of adenine nucleotides added to the 3′-end of the transcribed mRNA. It can comprise up to about 400 adenine nucleotides. In some embodiments the length of the 3′-poly(A) tail may be an essential element with respect to the stability of the individual mRNA.

In some embodiments the RNA (e.g., mRNA) vaccine may include one or more stabilizing elements. Stabilizing elements may include for instance a histone stem-loop. A stem-loop binding protein (SLBP), a 32 kDa protein has been identified. It is associated with the histone stem-loop at the 3′-end of the histone messages in both the nucleus and the cytoplasm. Its expression level is regulated by the cell cycle; it peaks during the S-phase, when histone mRNA levels are also elevated. The protein has been shown to be essential for efficient 3′-end processing of histone pre-mRNA by the U7 snRNP. SLBP continues to be associated with the stem-loop after processing, and then stimulates the translation of mature histone mRNAs into histone proteins in the cytoplasm. The RNA binding domain of SLBP is conserved through metazoa and protozoa; its binding to the histone stem-loop depends on the structure of the loop. The minimum binding site includes at least three nucleotides 5′ and two nucleotides 3′ relative to the stem-loop.

In some embodiments, the RNA (e.g., mRNA) vaccines include a coding region, at least one histone stem-loop, and optionally, a poly(A) sequence or polyadenylation signal. The poly(A) sequence or polyadenylation signal generally should enhance the expression level of the encoded protein. The encoded protein, in some embodiments, is not a histone protein, a reporter protein (e.g. Luciferase, GFP, EGFP, β-Galactosidase, EGFP), or a marker or selection protein (e.g. alpha-Globin, Galactokinase and Xanthine:guanine phosphoribosyl transferase (GPT)).

In some embodiments, the combination of a poly(A) sequence or polyadenylation signal and at least one histone stem-loop, even though both represent alternative mechanisms in nature, acts synergistically to increase the protein expression beyond the level observed with either of the individual elements. It has been found that the synergistic effect of the combination of poly(A) and at least one histone stem-loop does not depend on the order of the elements or the length of the poly(A) sequence.

In some embodiments, the RNA (e.g., mRNA) vaccine does not comprise a histone downstream element (HDE). “Histone downstream element” (HDE) includes a purine-rich polynucleotide stretch of approximately 15 to 20 nucleotides 3′ of naturally occurring stem-loops, representing the binding site for the U7 snRNA, which is involved in processing of histone pre-mRNA into mature histone mRNA. Ideally, the inventive nucleic acid does not include an intron.

In some embodiments, the RNA (e.g., mRNA) vaccine may or may not contain an enhancer and/or promoter sequence, which may be modified or unmodified or which may be activated or inactivated. In some embodiments, the histone stem-loop is generally derived from histone genes, and includes an intramolecular base pairing of two neighbored partially or entirely reverse complementary sequences separated by a spacer, including (e.g., consisting of) a short sequence, which forms the loop of the structure. The unpaired loop region is typically unable to base pair with either of the stem loop elements. It occurs more often in RNA, as is a key component of many RNA secondary structures, but may be present in single-stranded DNA as well. Stability of the stem-loop structure generally depends on the length, number of mismatches or bulges, and base composition of the paired region. In some embodiments, wobble base pairing (non-Watson-Crick base pairing) may result. In some embodiments, the at least one histone stem-loop sequence comprises a length of 15 to 45 nucleotides.

In other embodiments the RNA (e.g., mRNA) vaccine may have one or more AU-rich sequences removed. These sequences, sometimes referred to as AURES, are destabilizing sequences found in the 3′UTR. The AURES may be removed from the RNA (e.g., mRNA) vaccines. Alternatively the AURES may remain in the RNA (e.g., mRNA) vaccine.

Nanoparticle Formulations

In some embodiments, tropical disease RNA (e.g. mRNA) vaccines are formulated in a nanoparticle. In some embodiments, tropical disease RNA (e.g. mRNA) vaccines are formulated in a lipid nanoparticle. In some embodiments, tropical disease RNA (e.g. mRNA) vaccines are formulated in a lipid-polycation complex, referred to as a cationic lipid nanoparticle. As a non-limiting example, the polycation may include a cationic peptide or a polypeptide such as, but not limited to, polylysine, polyornithine and/or polyarginine. In some embodiments, tropical disease RNA (e.g., mRNA) vaccines are formulated in a lipid nanoparticle that includes a non-cationic lipid such as, but not limited to, cholesterol or dioleoyl phosphatidylethanolamine (DOPE).

A lipid nanoparticle formulation may be influenced by, but not limited to, the selection of the cationic lipid component, the degree of cationic lipid saturation, the nature of the PEGylation, ratio of all components and biophysical parameters such as size. In one example by Semple et al. (Nature Biotech. 2010 28:172-176), the lipid nanoparticle formulation is composed of 57.1% cationic lipid, 7.1% dipalmitoylphosphatidylcholine, 34.3% cholesterol, and 1.4% PEG-c-DMA. As another example, changing the composition of the cationic lipid can more effectively deliver siRNA to various antigen presenting cells (Basha et al. Mol Ther. 2011 19:2186-2200).

In some embodiments, lipid nanoparticle formulations may comprise 35 to 45% cationic lipid, 40% to 50% cationic lipid, 50% to 60% cationic lipid and/or 55% to 65% cationic lipid. In some embodiments, the ratio of lipid to RNA (e.g., mRNA) in lipid nanoparticles may be 5:1 to 20:1, 10:1 to 25:1, 15:1 to 30:1 and/or at least 30:1.

In some embodiments, the ratio of PEG in the lipid nanoparticle formulations may be increased or decreased and/or the carbon chain length of the PEG lipid may be modified from C14 to C18 to alter the pharmacokinetics and/or biodistribution of the lipid nanoparticle formulations. As a non-limiting example, lipid nanoparticle formulations may contain 0.5% to 3.0%, 1.0% to 3.5%, 1.5% to 4.0%, 2.0% to 4.5%, 2.5% to 5.0% and/or 3.0% to 6.0% of the lipid molar ratio of PEG-c-DOMG (R-3-[(ω-methoxy-poly(ethyleneglycol)2000)carbamoyl)]-1,2-dimyristyloxypropyl-3-amine) (also referred to herein as PEG-DOMG) as compared to the cationic lipid, DSPC and cholesterol. In some embodiments, the PEG-c-DOMG may be replaced with a PEG lipid such as, but not limited to, PEG-DSG (1,2-Distearoyl-sn-glycerol, methoxypolyethylene glycol), PEG-DMG (1,2-Dimyristoyl-sn-glycerol) and/or PEG-DPG (1,2-Dipalmitoyl-sn-glycerol, methoxypolyethylene glycol). The cationic lipid may be selected from any lipid known in the art such as, but not limited to, DLin-MC3-DMA, DLin-DMA, C12-200 and DLin-KC2-DMA.

In some embodiments, a tropical disease RNA (e.g. mRNA) vaccine formulation is a nanoparticle that comprises at least one lipid. The lipid may be selected from, but is not limited to, DLin-DMA, DLin-K-DMA, 98N12-5, C12-200, DLin-MC3-DMA, DLin-KC2-DMA, DODMA, PLGA, PEG, PEG-DMG, PEGylated lipids and amino alcohol lipids. In some embodiments, the lipid may be a cationic lipid such as, but not limited to, DLin-DMA, DLin-D-DMA, DLin-MC3-DMA, DLin-KC2-DMA, DODMA and amino alcohol lipids. The amino alcohol cationic lipid may be the lipids described in and/or made by the methods described in U.S. Patent Publication No. US20130150625, herein incorporated by reference in its entirety. As a non-limiting example, the cationic lipid may be 2-amino-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-2-{[(9Z,2Z)-octadeca-9,12-dien-1-yloxy]methyl}propan-1-ol (Compound 1 in US20130150625); 2-amino-3-[(9Z)-octadec-9-en-1-yloxy]-2-{[(9Z)-octadec-9-en-1-yloxy]methyl}propan-1-ol (Compound 2 in US20130150625); 2-amino-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-2-[(octyloxy)methyl]propan-1-ol (Compound 3 in US20130150625); and 2-(dimethylamino)-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-2-{[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]methyl}propan-1-ol (Compound 4 in US20130150625); or any pharmaceutically acceptable salt or stereoisomer thereof.

Lipid nanoparticle formulations typically comprise a lipid, in particular, an ionizable cationic lipid, for example, 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), or di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), and further comprise a neutral lipid, a sterol and a molecule capable of reducing particle aggregation, for example a PEG or PEG-modified lipid.

In some embodiments, a lipid nanoparticle formulation consists essentially of (i) at least one lipid selected from the group consisting of 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319); (ii) a neutral lipid selected from DSPC, DPPC, POPC, DOPE and SM; (iii) a sterol, e.g., cholesterol; and (iv) a PEG-lipid, e.g., PEG-DMG or PEG-cDMA, in a molar ratio of 20-60% cationic lipid:5-25% neutral lipid:25-55% sterol; 0.5-15% PEG-lipid.

In some embodiments, a lipid nanoparticle formulation includes 25% to 75% on a molar basis of a cationic lipid selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), e.g., 35% to 65%, 45% to 65%, 60%, 57.5%, 50% or 40% on a molar basis.

In some embodiments, a lipid nanoparticle formulation includes 0.5% to 15% on a molar basis of the neutral lipid, e.g., 3% to 12%, 5% to 10% or 15%, 10%, or 7.5% on a molar basis. Examples of neutral lipids include, without limitation, DSPC, POPC, DPPC, DOPE and SM. In some embodiments, the formulation includes 5% to 50% on a molar basis of the sterol (e.g., 15% to 45%, 20% to 40%, 40%, 38.5%, 35%, or 31% on a molar basis). A non-limiting example of a sterol is cholesterol. In some embodiments, a lipid nanoparticle formulation includes 0.5% to 20% on a molar basis of the PEG or PEG-modified lipid (e.g., 0.5% to 10%, 0.5% to 5%, 1.5%, 0.5%, 1.5%, 3.5%, or 5% on a molar basis). In some embodiments, a PEG or PEG modified lipid comprises a PEG molecule of an average molecular weight of 2,000 Da. In some embodiments, a PEG or PEG modified lipid comprises a PEG molecule of an average molecular weight of less than 2,000, for example around 1,500 Da, around 1,000 Da, or around 500 Da. Non-limiting examples of PEG-modified lipids include PEG-distearoyl glycerol (PEG-DMG) (also referred herein as PEG-C14 or C14-PEG), PEG-cDMA (further discussed in Reyes et al. J. Controlled Release, 107, 276-287 (2005) the contents of which are herein incorporated by reference in their entirety).

In some embodiments, lipid nanoparticle formulations include 25-75% of a cationic lipid selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), 0.5-15% of the neutral lipid, 5-50% of the sterol, and 0.5-20% of the PEG or PEG-modified lipid on a molar basis.

In some embodiments, lipid nanoparticle formulations include 35-65% of a cationic lipid selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), 3-12% of the neutral lipid, 15-45% of the sterol, and 0.5-10% of the PEG or PEG-modified lipid on a molar basis.

In some embodiments, lipid nanoparticle formulations include 45-65% of a cationic lipid selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), 5-10% of the neutral lipid, 25-40% of the sterol, and 0.5-10% of the PEG or PEG-modified lipid on a molar basis.

In some embodiments, lipid nanoparticle formulations include 60% of a cationic lipid selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), 7.5% of the neutral lipid, 31% of the sterol, and 1.5% of the PEG or PEG-modified lipid on a molar basis.

In some embodiments, lipid nanoparticle formulations include 50% of a cationic lipid selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), 10% of the neutral lipid, 38.5% of the sterol, and 1.5% of the PEG or PEG-modified lipid on a molar basis.

In some embodiments, lipid nanoparticle formulations include 50% of a cationic lipid selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), 10% of the neutral lipid, 35% of the sterol, 4.5% or 5% of the PEG or PEG-modified lipid, and 0.5% of the targeting lipid on a molar basis.

In some embodiments, lipid nanoparticle formulations include 40% of a cationic lipid selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), 15% of the neutral lipid, 40% of the sterol, and 5% of the PEG or PEG-modified lipid on a molar basis.

In some embodiments, lipid nanoparticle formulations include 57.2% of a cationic lipid selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), 7.1% of the neutral lipid, 34.3% of the sterol, and 1.4% of the PEG or PEG-modified lipid on a molar basis.

In some embodiments, lipid nanoparticle formulations include 57.5% of a cationic lipid selected from the PEG lipid is PEG-cDMA (PEG-cDMA is further discussed in Reyes et al. (J. Controlled Release, 107, 276-287 (2005), the contents of which are herein incorporated by reference in their entirety), 7.5% of the neutral lipid, 31.5% of the sterol, and 3.5% of the PEG or PEG-modified lipid on a molar basis.

In some embodiments, lipid nanoparticle formulations consist essentially of a lipid mixture in molar ratios of 20-70% cationic lipid: 5-45% neutral lipid: 20-55% cholesterol: 0.5-15% PEG-modified lipid. In some embodiments, lipid nanoparticle formulations consist essentially of a lipid mixture in a molar ratio of 20-60% cationic lipid:5-25% neutral lipid:25-55% cholesterol:0.5-15% PEG-modified lipid.

In some embodiments, the molar lipid ratio is 50/10/38.5/1.5 (mol % cationic lipid/neutral lipid, e.g., DSPC/Chol/PEG-modified lipid, e.g., PEG-DMG, PEG-DSG or PEG-DPG), 57.2/7.1134.3/1.4 (mol % cationic lipid/neutral lipid, e.g., DPPC/Chol/PEG-modified lipid, e.g., PEG-cDMA), 40/15/40/5 (mol % cationic lipid/neutral lipid, e.g., DSPC/Chol/PEG-modified lipid, e.g., PEG-DMG), 50/10/35/4.5/0.5 (mol % cationic lipid/neutral lipid, e.g., DSPC/Chol/PEG-modified lipid, e.g., PEG-DSG), 50/10/35/5 (cationic lipid/neutral lipid, e.g., DSPC/Chol/PEG-modified lipid, e.g., PEG-DMG), 40/10/40/10 (mol % cationic lipid/neutral lipid, e.g., DSPC/Chol/PEG-modified lipid, e.g., PEG-DMG or PEG-cDMA), 35/15/40/10 (mol % cationic lipid/neutral lipid, e.g., DSPC/Chol/PEG-modified lipid, e.g., PEG-DMG or PEG-cDMA) or 52/13/30/5 (mol % cationic lipid/neutral lipid, e.g., DSPC/Chol/PEG-modified lipid, e.g., PEG-DMG or PEG-cDMA).

Non-limiting examples of lipid nanoparticle compositions and methods of making them are described, for example, in Semple et al. (2010) Nat. Biotechnol. 28:172-176; Jayarama et al. (2012), Angew. Chem. Int. Ed., 51: 8529-8533; and Maier et al. (2013) Molecular Therapy 21, 1570-1578 (the contents of each of which are incorporated herein by reference in their entirety).

In some embodiments, lipid nanoparticle formulations may comprise a cationic lipid, a PEG lipid and a structural lipid and optionally comprise a non-cationic lipid. As a non-limiting example, a lipid nanoparticle may comprise 40-60% of cationic lipid, 5-15% of a non-cationic lipid, 1-2% of a PEG lipid and 30-50% of a structural lipid. As another non-limiting example, the lipid nanoparticle may comprise 50% cationic lipid, 10% non-cationic lipid, 1.5% PEG lipid and 38.5% structural lipid. As yet another non-limiting example, a lipid nanoparticle may comprise 55% cationic lipid, 10% non-cationic lipid, 2.5% PEG lipid and 32.5% structural lipid. In some embodiments, the cationic lipid may be any cationic lipid described herein such as, but not limited to, DLin-KC2-DMA, DLin-MC3-DMA and L319.

In some embodiments, the lipid nanoparticle formulations described herein may be 4 component lipid nanoparticles. The lipid nanoparticle may comprise a cationic lipid, a non-cationic lipid, a PEG lipid and a structural lipid. As a non-limiting example, the lipid nanoparticle may comprise 40-60% of cationic lipid, 5-15% of a non-cationic lipid, 1-2% of a PEG lipid and 30-50% of a structural lipid. As another non-limiting example, the lipid nanoparticle may comprise 50% cationic lipid, 10% non-cationic lipid, 1.5% PEG lipid and 38.5% structural lipid. As yet another non-limiting example, the lipid nanoparticle may comprise 55% cationic lipid, 10% non-cationic lipid, 2.5% PEG lipid and 32.5% structural lipid. In some embodiments, the cationic lipid may be any cationic lipid described herein such as, but not limited to, DLin-KC2-DMA, DLin-MC3-DMA and L319.

In some embodiments, the lipid nanoparticle formulations described herein may comprise a cationic lipid, a non-cationic lipid, a PEG lipid and a structural lipid. As a non-limiting example, the lipid nanoparticle comprises 50% of the cationic lipid DLin-KC2-DMA, 10% of the non-cationic lipid DSPC, 1.5% of the PEG lipid PEG-DOMG and 38.5% of the structural lipid cholesterol. As a non-limiting example, the lipid nanoparticle comprises 50% of the cationic lipid DLin-MC3-DMA, 10% of the non-cationic lipid DSPC, 1.5% of the PEG lipid PEG-DOMG and 38.5% of the structural lipid cholesterol. As a non-limiting example, the lipid nanoparticle comprises 50% of the cationic lipid DLin-MC3-DMA, 10% of the non-cationic lipid DSPC, 1.5% of the PEG lipid PEG-DMG and 38.5% of the structural lipid cholesterol. As yet another non-limiting example, the lipid nanoparticle comprises 55% of the cationic lipid L319, 10% of the non-cationic lipid DSPC, 2.5% of the PEG lipid PEG-DMG and 32.5% of the structural lipid cholesterol.

Relative amounts of the active ingredient, the pharmaceutically acceptable excipient, and/or any additional ingredients in a vaccine composition may vary, depending upon the identity, size, and/or condition of the subject being treated and further depending upon the route by which the composition is to be administered. For example, the composition may comprise between 0.1% and 99% (w/w) of the active ingredient. By way of example, the composition may comprise between 0.1% and 100%, e.g., between 0.5% and 50%, between 1-30%, between 5-80%, at least 80% (w/w) active ingredient.

In some embodiments, the tropical disease RNA (e.g. mRNA) vaccine composition may comprise the polynucleotide described herein, formulated in a lipid nanoparticle comprising MC3, Cholesterol, DSPC and PEG2000-DMG, the buffer trisodium citrate, sucrose and water for injection. As a non-limiting example, the composition comprises: 2.0 mg/mL of drug substance, 21.8 mg/mL of MC3, 10.1 mg/mL of cholesterol, 5.4 mg/mL of DSPC, 2.7 mg/mL of PEG2000-DMG, 5.16 mg/mL of trisodium citrate, 71 mg/mL of sucrose and 1.0 mL of water for injection.

In some embodiments, a nanoparticle (e.g., a lipid nanoparticle) has a mean diameter of 10-500 nm, 20-400 nm, 30-300 nm, or 40-200 nm. In some embodiments, a nanoparticle (e.g., a lipid nanoparticle) has a mean diameter of 50-150 nm, 50-200 nm, 80-100 nm or 80-200 nm.

Liposomes, Lipoplexes, and Lipid Nanoparticles

The RNA (e.g., mRNA) vaccines of the disclosure can be formulated using one or more liposomes, lipoplexes, or lipid nanoparticles. In some embodiments, pharmaceutical compositions of RNA (e.g., mRNA) vaccines include liposomes. Liposomes are artificially-prepared vesicles which may primarily be composed of a lipid bilayer and may be used as a delivery vehicle for the administration of nutrients and pharmaceutical formulations. Liposomes can be of different sizes such as, but not limited to, a multilamellar vesicle (MLV) which may be hundreds of nanometers in diameter and may contain a series of concentric bilayers separated by narrow aqueous compartments, a small unicellular vesicle (SUV) which may be smaller than 50 nm in diameter, and a large unilamellar vesicle (LUV) which may be between 50 and 500 nm in diameter. Liposome design may include, but is not limited to, opsonins or ligands in order to improve the attachment of liposomes to unhealthy tissue or to activate events such as, but not limited to, endocytosis. Liposomes may contain a low or a high pH in order to improve the delivery of the pharmaceutical formulations.

The formation of liposomes may depend on the physicochemical characteristics such as, but not limited to, the pharmaceutical formulation entrapped and the liposomal ingredients, the nature of the medium in which the lipid vesicles are dispersed, the effective concentration of the entrapped substance and its potential toxicity, any additional processes involved during the application and/or delivery of the vesicles, the optimization size, polydispersity and the shelf-life of the vesicles for the intended application, and the batch-to-batch reproducibility and possibility of large-scale production of safe and efficient liposomal products.

In some embodiments, pharmaceutical compositions described herein may include, without limitation, liposomes such as those formed from 1,2-dioleyloxy-N,N-dimethylaminopropane (DODMA) liposomes, DiLa2 liposomes from Marina Biotech (Bothell, Wash.), 1,2-dilinoleyloxy-3-dimethylaminopropane (DLin-DMA), 2,2-dilinoleyl-4-(2-dimethylaminoethyl)[1,3]-dioxolane (DLin-KC2-DMA), and MC3 (US20100324120; herein incorporated by reference in its entirety) and liposomes which may deliver small molecule drugs such as, but not limited to, DOXIL® from Janssen Biotech, Inc. (Horsham, Pa.).

In some embodiments, pharmaceutical compositions described herein may include, without limitation, liposomes such as those formed from the synthesis of stabilized plasmid-lipid particles (SPLP) or stabilized nucleic acid lipid particle (SNALP) that have been previously described and shown to be suitable for oligonucleotide delivery in vitro and in vivo (see Wheeler et al. Gene Therapy. 1999 6:271-281; Zhang et al. Gene Therapy. 1999 6:1438-1447; Jeffs et al. Pharm Res. 2005 22:362-372; Morrissey et al., Nat Biotechnol. 2005 2:1002-1007; Zimmermann et al., Nature. 2006 441:111-114; Heyes et al. J Contr Rel. 2005 107:276-287; Semple et al. Nature Biotech. 2010 28:172-176; Judge et al. J Clin Invest. 2009 119:661-673; deFougerolles Hum Gene Ther. 2008 19:125-132; U.S. Patent Publication No US20130122104; all of which are incorporated herein in their entireties). The original manufacture method by Wheeler et al. was a detergent dialysis method, which was later improved by Jeffs et al. and is referred to as the spontaneous vesicle formation method. The liposome formulations are composed of 3 to 4 lipid components in addition to the polynucleotide. As an example a liposome can contain, but is not limited to, 55% cholesterol, 20% disteroylphosphatidyl choline (DSPC), 10% PEG-S-DSG, and 15% 1,2-dioleyloxy-N,N-dimethylaminopropane (DODMA), as described by Jeffs et al. As another example, certain liposome formulations may contain, but are not limited to, 48% cholesterol, 20% DSPC, 2% PEG-c-DMA, and 30% cationic lipid, where the cationic lipid can be 1,2-distearloxy-N,N-dimethylaminopropane (DSDMA), DODMA, DLin-DMA, or 1,2-dilinolenyloxy-3-dimethylaminopropane (DLenDMA), as described by Heyes et al.

In some embodiments, liposome formulations may comprise from about 25.0% cholesterol to about 40.0% cholesterol, from about 30.0% cholesterol to about 45.0% cholesterol, from about 35.0% cholesterol to about 50.0% cholesterol and/or from about 48.5% cholesterol to about 60% cholesterol. In some embodiments, formulations may comprise a percentage of cholesterol selected from the group consisting of 28.5%, 31.5%, 33.5%, 36.5%, 37.0%, 38.5%, 39.0% and 43.5%. In some embodiments, formulations may comprise from about 5.0% to about 10.0% DSPC and/or from about 7.0% to about 15.0% DSPC.

In some embodiments, the RNA (e.g., mRNA) vaccine pharmaceutical compositions may be formulated in liposomes such as, but not limited to, DiLa2 liposomes (Marina Biotech, Bothell, Wash.), SMARTICLES® (Marina Biotech, Bothell, Wash.), neutral DOPC (1,2-dioleoyl-sn-glycero-3-phosphocholine) based liposomes (e.g., siRNA delivery for ovarian cancer (Landen et al. Cancer Biology & Therapy 2006 5(12)1708-1713); herein incorporated by reference in its entirety) and hyaluronan-coated liposomes (Quiet Therapeutics, Israel).

In some embodiments, the cationic lipid may be a low molecular weight cationic lipid such as those described in U.S. Patent Application No. 20130090372, the contents of which are herein incorporated by reference in their entirety.

In some embodiments, the RNA (e.g., mRNA) vaccines may be formulated in a lipid vesicle, which may have crosslinks between functionalized lipid bilayers.

In some embodiments, the RNA (e.g., mRNA) vaccines may be formulated in a lipid-polycation complex. The formation of the lipid-polycation complex may be accomplished by methods known in the art and/or as described in U.S. Pub. No. 20120178702, herein incorporated by reference in its entirety. As a non-limiting example, the polycation may include a cationic peptide or a polypeptide such as, but not limited to, polylysine, polyornithine and/or polyarginine. In some embodiments, the RNA (e.g., mRNA) vaccines may be formulated in a lipid-polycation complex, which may further include a non-cationic lipid such as, but not limited to, cholesterol or dioleoyl phosphatidylethanolamine (DOPE).

In some embodiments, the ratio of PEG in the lipid nanoparticle (LNP) formulations may be increased or decreased and/or the carbon chain length of the PEG lipid may be modified from C14 to C18 to alter the pharmacokinetics and/or biodistribution of the LNP formulations. As a non-limiting example, LNP formulations may contain from about 0.5% to about 3.0%, from about 1.0% to about 3.5%, from about 1.5% to about 4.0%, from about 2.0% to about 4.5%, from about 2.5% to about 5.0% and/or from about 3.0% to about 6.0% of the lipid molar ratio of PEG-c-DOMG (R-3-[(ω-methoxy-poly(ethyleneglycol)2000)carbamoyl)]-1,2-dimyristyloxypropyl-3-amine) (also referred to herein as PEG-DOMG) as compared to the cationic lipid, DSPC and cholesterol. In some embodiments, the PEG-c-DOMG may be replaced with a PEG lipid such as, but not limited to, PEG-DSG (1,2-Distearoyl-sn-glycerol, methoxypolyethylene glycol), PEG-DMG (1,2-Dimyristoyl-sn-glycerol) and/or PEG-DPG (1,2-Dipalmitoyl-sn-glycerol, methoxypolyethylene glycol). The cationic lipid may be selected from any lipid known in the art such as, but not limited to, DLin-MC3-DMA, DLin-DMA, C12-200 and DLin-KC2-DMA.

In some embodiments, the RNA (e.g., mRNA) vaccines may be formulated in a lipid nanoparticle.

In some embodiments, the RNA (e.g., mRNA) vaccine formulation comprising the polynucleotide is a nanoparticle which may comprise at least one lipid. The lipid may be selected from, but is not limited to, DLin-DMA, DLin-K-DMA, 98N12-5, C12-200, DLin-MC3-DMA, DLin-KC2-DMA, DODMA, PLGA, PEG, PEG-DMG, PEGylated lipids and amino alcohol lipids. In another aspect, the lipid may be a cationic lipid such as, but not limited to, DLin-DMA, DLin-D-DMA, DLin-MC3-DMA, DLin-KC2-DMA, DODMA and amino alcohol lipids. The amino alcohol cationic lipid may be the lipids described in and/or made by the methods described in U.S. Patent Publication No. US20130150625, herein incorporated by reference in its entirety. As a non-limiting example, the cationic lipid may be 2-amino-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-2-{[(9Z,2Z)-octadeca-9,12-dien-1-yloxy]methyl}propan-1-ol (Compound 1 in US20130150625); 2-amino-3-[(9Z)-octadec-9-en-1-yloxy]-2-{[(9Z)-octadec-9-en-1-yloxy]methyl}propan-1-ol (Compound 2 in US20130150625); 2-amino-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-2-[(octyloxy)methyl]propan-1-ol (Compound 3 in US20130150625); and 2-(dimethylamino)-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]-2-{[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]methyl}propan-1-ol (Compound 4 in US20130150625); or any pharmaceutically acceptable salt or stereoisomer thereof.

Lipid nanoparticle formulations typically comprise a lipid, in particular, an ionizable cationic lipid, for example, 2,2-dilinoleyl-4-dimethylaminoethyl[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), or di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), and further comprise a neutral lipid, a sterol and a molecule capable of reducing particle aggregation, for example a PEG or PEG-modified lipid.

In some embodiments, the lipid nanoparticle formulation consists essentially of (i) at least one lipid selected from the group consisting of 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319); (ii) a neutral lipid selected from DSPC, DPPC, POPC, DOPE and SM; (iii) a sterol, e.g., cholesterol; and (iv) a PEG-lipid, e.g., PEG-DMG or PEG-cDMA, in a molar ratio of about 20-60% cationic lipid:5-25% neutral lipid:25-55% sterol; 0.5-15% PEG-lipid.

In some embodiments, the formulation includes from about 25% to about 75% on a molar basis of a cationic lipid selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), e.g., from about 35 to about 65%, from about 45 to about 65%, about 60%, about 57.5%, about 50% or about 40% on a molar basis.

In some embodiments, the formulation includes from about 0.5% to about 15% on a molar basis of the neutral lipid e.g., from about 3 to about 12%, from about 5 to about 10% or about 15%, about 10%, or about 7.5% on a molar basis. Examples of neutral lipids include, but are not limited to, DSPC, POPC, DPPC, DOPE and SM. In some embodiments, the formulation includes from about 5% to about 50% on a molar basis of the sterol (e.g., about 15 to about 45%, about 20 to about 40%, about 40%, about 38.5%, about 35%, or about 31% on a molar basis. An exemplary sterol is cholesterol. In some embodiments, the formulation includes from about 0.5% to about 20% on a molar basis of the PEG or PEG-modified lipid (e.g., about 0.5 to about 10%, about 0.5 to about 5%, about 1.5%, about 0.5%, about 1.5%, about 3.5%, or about 5% on a molar basis). In some embodiments, the PEG or PEG modified lipid comprises a PEG molecule of an average molecular weight of 2,000 Da. In other embodiments, the PEG or PEG modified lipid comprises a PEG molecule of an average molecular weight of less than 2,000, for example around 1,500 Da, around 1,000 Da, or around 500 Da. Examples of PEG-modified lipids include, but are not limited to, PEG-distearoyl glycerol (PEG-DMG) (also referred herein as PEG-C14 or C14-PEG), PEG-cDMA (further discussed in Reyes et al. J. Controlled Release, 107, 276-287 (2005) the contents of which are herein incorporated by reference in their entirety)

In some embodiments, the formulations of the present disclosure include 25-75% of a cationic lipid selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), 0.5-15% of the neutral lipid, 5-50% of the sterol, and 0.5-20% of the PEG or PEG-modified lipid on a molar basis.

In some embodiments, the formulations of the present disclosure include 35-65% of a cationic lipid selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), 3-12% of the neutral lipid, 15-45% of the sterol, and 0.5-10% of the PEG or PEG-modified lipid on a molar basis.

In some embodiments, the formulations of the present disclosure include 45-65% of a cationic lipid selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), 5-10% of the neutral lipid, 25-40% of the sterol, and 0.5-10% of the PEG or PEG-modified lipid on a molar basis.

In some embodiments, the formulations of the present disclosure include about 60% of a cationic lipid selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), about 7.5% of the neutral lipid, about 31% of the sterol, and about 1.5% of the PEG or PEG-modified lipid on a molar basis.

In some embodiments, the formulations of the present disclosure include about 50% of a cationic lipid selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), about 10% of the neutral lipid, about 38.5% of the sterol, and about 1.5% of the PEG or PEG-modified lipid on a molar basis.

In some embodiments, the formulations of the present disclosure include about 50% of a cationic lipid selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), about 10% of the neutral lipid, about 35% of the sterol, about 4.5% or about 5% of the PEG or PEG-modified lipid, and about 0.5% of the targeting lipid on a molar basis.

In some embodiments, the formulations of the present disclosure include about 40% of a cationic lipid selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), about 15% of the neutral lipid, about 40% of the sterol, and about 5% of the PEG or PEG-modified lipid on a molar basis.

In some embodiments, the formulations of the present disclosure include about 57.2% of a cationic lipid selected from 2,2-dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), dilinoleyl-methyl-4-dimethylaminobutyrate (DLin-MC3-DMA), and di((Z)-non-2-en-1-yl) 9-((4-(dimethylamino)butanoyl)oxy)heptadecanedioate (L319), about 7.1% of the neutral lipid, about 34.3% of the sterol, and about 1.4% of the PEG or PEG-modified lipid on a molar basis.

In some embodiments, the formulations of the present disclosure include about 57.5% of a cationic lipid selected from the PEG lipid is PEG-cDMA (PEG-cDMA is further discussed in Reyes et al. (J. Controlled Release, 107, 276-287 (2005), the contents of which are herein incorporated by reference in their entirety), about 7.5% of the neutral lipid, about 31.5% of the sterol, and about 3.5% of the PEG or PEG-modified lipid on a molar basis.

In some embodiments, lipid nanoparticle formulation consists essentially of a lipid mixture in molar ratios of about 20-70% cationic lipid:5-45% neutral lipid:20-55% cholesterol:0.5-15% PEG-modified lipid; more preferably in a molar ratio of about 20-60% cationic lipid:5-25% neutral lipid:25-55% cholesterol:0.5-15% PEG-modified lipid.

In some embodiments, the molar lipid ratio is approximately 50/10/38.5/1.5 (mol % cationic lipid/neutral lipid, e.g., DSPC/Chol/PEG-modified lipid, e.g., PEG-DMG, PEG-DSG or PEG-DPG), 57.2/7.1134.3/1.4 (mol % cationic lipid/neutral lipid, e.g., DPPC/Chol/PEG-modified lipid, e.g., PEG-cDMA), 40/15/40/5 (mol % cationic lipid/neutral lipid, e.g., DSPC/Chol/PEG-modified lipid, e.g., PEG-DMG), 50/10/35/4.5/0.5 (mol % cationic lipid/neutral lipid, e.g., DSPC/Chol/PEG-modified lipid, e.g., PEG-DSG), 50/10/35/5 (cationic lipid/neutral lipid, e.g., DSPC/Chol/PEG-modified lipid, e.g., PEG-DMG), 40/10/40/10 (mol % cationic lipid/neutral lipid, e.g., DSPC/Chol/PEG-modified lipid, e.g., PEG-DMG or PEG-cDMA), 35/15/40/10 (mol % cationic lipid/neutral lipid, e.g., DSPC/Chol/PEG-modified lipid, e.g., PEG-DMG or PEG-cDMA) or 52/13/30/5 (mol % cationic lipid/neutral lipid, e.g., DSPC/Chol/PEG-modified lipid, e.g., PEG-DMG or PEG-cDMA).

Examples of lipid nanoparticle compositions and methods of making same are described, for example, in Semple et al. (2010) Nat. Biotechnol. 28:172-176; Jayarama et al. (2012), Angew. Chem. Int. Ed., 51: 8529-8533; and Maier et al. (2013) Molecular Therapy 21, 1570-1578 (the contents of each of which are incorporated herein by reference in their entirety).

In some embodiments, the LNP formulations of the RNA (e.g., mRNA) vaccines may contain PEG-c-DOMG at 3% lipid molar ratio. In some embodiments, the LNP formulations of the RNA (e.g., mRNA) vaccines may contain PEG-c-DOMG at 1.5% lipid molar ratio.

In some embodiments, the pharmaceutical compositions of the RNA (e.g., mRNA) vaccines may include at least one of the PEGylated lipids described in International Publication No. WO2012099755, the contents of which are herein incorporated by reference in their entirety.

In some embodiments, the LNP formulation may contain PEG-DMG 2000 (1,2-dimyristoyl-sn-glycero-3-phophoethanolamine-N-[methoxy(polyethylene glycol)-2000). In some embodiments, the LNP formulation may contain PEG-DMG 2000, a cationic lipid known in the art and at least one other component. In some embodiments, the LNP formulation may contain PEG-DMG 2000, a cationic lipid known in the art, DSPC and cholesterol. As a non-limiting example, the LNP formulation may contain PEG-DMG 2000, DLin-DMA, DSPC and cholesterol. As another non-limiting example the LNP formulation may contain PEG-DMG 2000, DLin-DMA, DSPC and cholesterol in a molar ratio of 2:40:10:48 (see e.g., Geall et al., Nonviral delivery of self-amplifying RNA (e.g., mRNA) vaccines, PNAS 2012; PMID: 22908294, the contents of each of which are herein incorporated by reference in their entirety).

The lipid nanoparticles described herein may be made in a sterile environment.

In some embodiments, the LNP formulation may be formulated in a nanoparticle such as a nucleic acid-lipid particle. As a non-limiting example, the lipid particle may comprise one or more active agents or therapeutic agents; one or more cationic lipids comprising from about 50 mol % to about 85 mol % of the total lipid present in the particle; one or more non-cationic lipids comprising from about 13 mol % to about 49.5 mol % of the total lipid present in the particle; and one or more conjugated lipids that inhibit aggregation of particles comprising from about 0.5 mol % to about 2 mol % of the total lipid present in the particle.

The nanoparticle formulations may comprise a phosphate conjugate. The phosphate conjugate may increase in vivo circulation times and/or increase the targeted delivery of the nanoparticle. As a non-limiting example, the phosphate conjugates may include a compound of any one of the formulas described in International Application No. WO2013033438, the contents of which are herein incorporated by reference in its entirety.

The nanoparticle formulation may comprise a polymer conjugate. The polymer conjugate may be a water-soluble conjugate. The polymer conjugate may have a structure as described in U.S. Patent Application No. 20130059360, the contents of which are herein incorporated by reference in its entirety. In some embodiments, polymer conjugates with the polynucleotides of the present disclosure may be made using the methods and/or segmented polymeric reagents described in U.S. Patent Application No. 20130072709, the contents of which are herein incorporated by reference in its entirety. In some embodiments, the polymer conjugate may have pendant side groups comprising ring moieties such as, but not limited to, the polymer conjugates described in U.S. Patent Publication No. US20130196948, the contents which are herein incorporated by reference in its entirety.

The nanoparticle formulations may comprise a conjugate to enhance the delivery of nanoparticles of the present disclosure in a subject. Further, the conjugate may inhibit phagocytic clearance of the nanoparticles in a subject. In one aspect, the conjugate may be a “self” peptide designed from the human membrane protein CD47 (e.g., the “self” particles described by Rodriguez et al. (Science 2013 339, 971-975), herein incorporated by reference in its entirety). As shown by Rodriguez et al., the self peptides delayed macrophage-mediated clearance of nanoparticles which enhanced delivery of the nanoparticles. In another aspect, the conjugate may be the membrane protein CD47 (e.g., see Rodriguez et al. Science 2013 339, 971-975, herein incorporated by reference in its entirety). Rodriguez et al. showed that, similarly to “self” peptides, CD47 can increase the circulating particle ratio in a subject as compared to scrambled peptides and PEG coated nanoparticles.

In some embodiments, the RNA (e.g., mRNA) vaccines of the present disclosure are formulated in nanoparticles which comprise a conjugate to enhance the delivery of the nanoparticles of the present disclosure in a subject. The conjugate may be the CD47 membrane or the conjugate may be derived from the CD47 membrane protein, such as the “self” peptide described previously. In some embodiments, the nanoparticle may comprise PEG and a conjugate of CD47 or a derivative thereof. In some embodiments, the nanoparticle may comprise both the “self” peptide described above and the membrane protein CD47.

In some embodiments, a “self” peptide and/or CD47 protein may be conjugated to a virus-like particle or pseudovirion, as described herein for delivery of the RNA (e.g., mRNA) vaccines of the present disclosure.

In some embodiments, RNA (e.g., mRNA) vaccine pharmaceutical compositions comprising the polynucleotides of the present disclosure and a conjugate that may have a degradable linkage. Non-limiting examples of conjugates include an aromatic moiety comprising an ionizable hydrogen atom, a spacer moiety, and a water-soluble polymer. As a non-limiting example, pharmaceutical compositions comprising a conjugate with a degradable linkage and methods for delivering such pharmaceutical compositions are described in U.S. Patent Publication No. US20130184443, the contents of which are herein incorporated by reference in their entirety.

The nanoparticle formulations may be a carbohydrate nanoparticle comprising a carbohydrate carrier and a RNA (e.g., mRNA) vaccine. As a non-limiting example, the carbohydrate carrier may include, but is not limited to, an anhydride-modified phytoglycogen or glycogen-type material, phytoglycogen octenyl succinate, phytoglycogen beta-dextrin, anhydride-modified phytoglycogen beta-dextrin. (See e.g., International Publication No. WO2012109121; the contents of which are herein incorporated by reference in their entirety).

Nanoparticle formulations of the present disclosure may be coated with a surfactant or polymer in order to improve the delivery of the particle. In some embodiments, the nanoparticle may be coated with a hydrophilic coating such as, but not limited to, PEG coatings and/or coatings that have a neutral surface charge. The hydrophilic coatings may help to deliver nanoparticles with larger payloads such as, but not limited to, RNA (e.g., mRNA) vaccines within the central nervous system. As a non-limiting example nanoparticles comprising a hydrophilic coating and methods of making such nanoparticles are described in U.S. Patent Publication No. US20130183244, the contents of which are herein incorporated by reference in their entirety.

In some embodiments, the lipid nanoparticles of the present disclosure may be hydrophilic polymer particles. Non-limiting examples of hydrophilic polymer particles and methods of making hydrophilic polymer particles are described in U.S. Patent Publication No. US20130210991, the contents of which are herein incorporated by reference in their entirety.

In some embodiments, the lipid nanoparticles of the present disclosure may be hydrophobic polymer particles.

Lipid nanoparticle formulations may be improved by replacing the cationic lipid with a biodegradable cationic lipid which is known as a rapidly eliminated lipid nanoparticle (reLNP). Ionizable cationic lipids, such as, but not limited to, DLinDMA, DLin-KC2-DMA, and DLin-MC3-DMA, have been shown to accumulate in plasma and tissues over time and may be a potential source of toxicity. The rapid metabolism of the rapidly eliminated lipids can improve the tolerability and therapeutic index of the lipid nanoparticles by an order of magnitude from a 1 mg/kg dose to a 10 mg/kg dose in rat. Inclusion of an enzymatically degraded ester linkage can improve the degradation and metabolism profile of the cationic component, while still maintaining the activity of the reLNP formulation. The ester linkage can be internally located within the lipid chain or it may be terminally located at the terminal end of the lipid chain. The internal ester linkage may replace any carbon in the lipid chain.

In some embodiments, the internal ester linkage may be located on either side of the saturated carbon.

In some embodiments, an immune response may be elicited by delivering a lipid nanoparticle which may include a nanospecies, a polymer and an immunogen. (U.S. Publication No. 20120189700 and International Publication No. WO2012099805; each of which is herein incorporated by reference in their entirety). The polymer may encapsulate the nanospecies or partially encapsulate the nanospecies. The immunogen may be a recombinant protein, a modified RNA and/or a polynucleotide described herein. In some embodiments, the lipid nanoparticle may be formulated for use in a vaccine such as, but not limited to, against a pathogen.

Lipid nanoparticles may be engineered to alter the surface properties of particles so the lipid nanoparticles may penetrate the mucosal barrier. Mucus is located on mucosal tissue such as, but not limited to, oral (e.g., the buccal and esophageal membranes and tonsil tissue), ophthalmic, gastrointestinal (e.g., stomach, small intestine, large intestine, colon, rectum), nasal, respiratory (e.g., nasal, pharyngeal, tracheal and bronchial membranes), genital (e.g., vaginal, cervical and urethral membranes). Nanoparticles larger than 10-200 nm which are preferred for higher drug encapsulation efficiency and the ability to provide the sustained delivery of a wide array of drugs have been thought to be too large to rapidly diffuse through mucosal barriers. Mucus is continuously secreted, shed, discarded or digested and recycled so most of the trapped particles may be removed from the mucosa tissue within seconds or within a few hours. Large polymeric nanoparticles (200 nm-500 nm in diameter) which have been coated densely with a low molecular weight polyethylene glycol (PEG) diffused through mucus only 4- to 6-fold lower than the same particles diffusing in water (Lai et al. PNAS 2007 104:1482-487; Lai et al. Adv Drug Deliv Rev. 2009 61: 158-171; each of which is herein incorporated by reference in its entirety). The transport of nanoparticles may be determined using rates of permeation and/or fluorescent microscopy techniques including, but not limited to, fluorescence recovery after photobleaching (FRAP) and high resolution multiple particle tracking (MPT). As a non-limiting example, compositions which can penetrate a mucosal barrier may be made as described in U.S. Pat. No. 8,241,670 or International Patent Publication No. WO2013110028, the contents of each of which are herein incorporated by reference in its entirety.

The lipid nanoparticle engineered to penetrate mucus may comprise a polymeric material (i.e. a polymeric core) and/or a polymer-vitamin conjugate and/or a tri-block co-polymer. The polymeric material may include, but is not limited to, polyamines, polyethers, polyamides, polyesters, polycarbamates, polyureas, polycarbonates, poly(styrenes), polyimides, polysulfones, polyurethanes, polyacetylenes, polyethylenes, polyethyeneimines, polyisocyanates, polyacrylates, polymethacrylates, polyacrylonitriles, and polyarylates. The polymeric material may be biodegradable and/or biocompatible. Non-limiting examples of biocompatible polymers are described in International Patent Publication No. WO2013116804, the contents of which are herein incorporated by reference in their entirety. The polymeric material may additionally be irradiated. As a non-limiting example, the polymeric material may be gamma irradiated (see e.g., International App. No. WO201282165, herein incorporated by reference in its entirety). Non-limiting examples of specific polymers include poly(caprolactone) (PCL), ethylene vinyl acetate polymer (EVA), poly(lactic acid) (PLA), poly(L-lactic acid) (PLLA), poly(glycolic acid) (PGA), poly(lactic acid-co-glycolic acid) (PLGA), poly(L-lactic acid-co-glycolic acid) (PLLGA), poly(D,L-lactide) (PDLA), poly(L-lactide) (PLLA), poly(D,L-lactide-co-caprolactone), poly(D,L-lactide-co-caprolactone-co-glycolide), poly(D,L-lactide-co-PEO-co-D,L-lactide), poly(D,L-lactide-co-PPO-co-D,L-lactide), polyalkyl cyanoacralate, polyurethane, poly-L-lysine (PLL), hydroxypropyl methacrylate (HPMA), polyethyleneglycol, poly-L-glutamic acid, poly(hydroxy acids), polyanhydrides, polyorthoesters, poly(ester amides), polyamides, poly(ester ethers), polycarbonates, polyalkylenes such as polyethylene and polypropylene, polyalkylene glycols such as poly(ethylene glycol) (PEG), polyalkylene oxides (PEO), polyalkylene terephthalates such as poly(ethylene terephthalate), polyvinyl alcohols (PVA), polyvinyl ethers, polyvinyl esters such as poly(vinyl acetate), polyvinyl halides such as poly(vinyl chloride) (PVC), polyvinylpyrrolidone, polysiloxanes, polystyrene (PS), polyurethanes, derivatized celluloses such as alkyl celluloses, hydroxyalkyl celluloses, cellulose ethers, cellulose esters, nitro celluloses, hydroxypropylcellulose, carboxymethylcellulose, polymers of acrylic acids, such as poly(methyl(meth)acrylate) (PMMA), poly(ethyl(meth)acrylate), poly(butyl(meth)acrylate), poly(isobutyl(meth)acrylate), poly(hexyl(meth)acrylate), poly(isodecyl(meth)acrylate), poly(lauryl(meth)acrylate), poly(phenyl(meth)acrylate), poly(methyl acrylate), poly(isopropyl acrylate), poly(isobutyl acrylate), poly(octadecyl acrylate) and copolymers and mixtures thereof, polydioxanone and its copolymers, polyhydroxyalkanoates, polypropylene fumarate, polyoxymethylene, poloxamers, poly(ortho)esters, poly(butyric acid), poly(valeric acid), poly(lactide-co-caprolactone), PEG-PLGA-PEG and trimethylene carbonate, polyvinylpyrrolidone. The lipid nanoparticle may be coated or associated with a co-polymer such as, but not limited to, a block co-polymer (such as a branched polyether-polyamide block copolymer described in International Publication No. WO2013012476, herein incorporated by reference in its entirety), and (poly(ethylene glycol))-(poly(propylene oxide))-(poly(ethylene glycol)) triblock copolymer (see e.g., U.S. Publication 20120121718 and U.S. Publication 20100003337 and U.S. Pat. No. 8,263,665, the contents of each of which is herein incorporated by reference in their entirety). The co-polymer may be a polymer that is generally regarded as safe (GRAS) and the formation of the lipid nanoparticle may be in such a way that no new chemical entities are created. For example, the lipid nanoparticle may comprise poloxamers coating PLGA nanoparticles without forming new chemical entities which are still able to rapidly penetrate human mucus (Yang et al. Angew. Chem. Int. Ed. 2011 50:2597-2600; the contents of which are herein incorporated by reference in their entirety). A non-limiting scalable method to produce nanoparticles which can penetrate human mucus is described by Xu et al. (see, e.g., J Control Release 2013, 170:279-86; the contents of which are herein incorporated by reference in their entirety).

The vitamin of the polymer-vitamin conjugate may be vitamin E. The vitamin portion of the conjugate may be substituted with other suitable components such as, but not limited to, vitamin A, vitamin E, other vitamins, cholesterol, a hydrophobic moiety, or a hydrophobic component of other surfactants (e.g., sterol chains, fatty acids, hydrocarbon chains and alkylene oxide chains).

The lipid nanoparticle engineered to penetrate mucus may include surface altering agents such as, but not limited to, polynucleotides, anionic proteins (e.g., bovine serum albumin), surfactants (e.g., cationic surfactants such as for example dimethyldioctadecylammonium bromide), sugars or sugar derivatives (e.g., cyclodextrin), nucleic acids, polymers (e.g., heparin, polyethylene glycol and poloxamer), mucolytic agents (e.g., N-acetylcysteine, mugwort, bromelain, papain, clerodendrum, acetylcysteine, bromhexine, carbocisteine, eprazinone, mesna, ambroxol, sobrerol, domiodol, letosteine, stepronin, tiopronin, gelsolin, thymosin β4 dornase alfa, neltenexine, erdosteine) and various DNases including rhDNase. The surface altering agent may be embedded or enmeshed in the particle's surface or disposed (e.g., by coating, adsorption, covalent linkage, or other process) on the surface of the lipid nanoparticle. (see e.g., U.S. Publication 20100215580 and U.S. Publication 20080166414 and US20130164343; the contents of each of which are herein incorporated by reference in their entirety).

In some embodiments, the mucus penetrating lipid nanoparticles may comprise at least one polynucleotide described herein. The polynucleotide may be encapsulated in the lipid nanoparticle and/or disposed on the surface of the particle. The polynucleotide may be covalently coupled to the lipid nanoparticle. Formulations of mucus penetrating lipid nanoparticles may comprise a plurality of nanoparticles. Further, the formulations may contain particles which may interact with the mucus and alter the structural and/or adhesive properties of the surrounding mucus to decrease mucoadhesion, which may increase the delivery of the mucus penetrating lipid nanoparticles to the mucosal tissue.

In some embodiments, the mucus penetrating lipid nanoparticles may be a hypotonic formulation comprising a mucosal penetration enhancing coating. The formulation may be hypotonic for the epithelium to which it is being delivered. Non-limiting examples of hypotonic formulations may be found in International Patent Publication No. WO2013110028, the contents of which are herein incorporated by reference in their entirety.

In some embodiments, in order to enhance the delivery through the mucosal barrier the RNA (e.g., mRNA) vaccine formulation may comprise or be a hypotonic solution. Hypotonic solutions were found to increase the rate at which mucoinert particles such as, but not limited to, mucus-penetrating particles, were able to reach the vaginal epithelial surface (see e.g., Ensign et al. Biomaterials 2013 34(28):6922-9, the contents of which are herein incorporated by reference in their entirety).

In some embodiments, the RNA (e.g., mRNA) vaccine is formulated as a lipoplex, such as, without limitation, the ATUPLEX™® system, the DACC system, the DBTC system and other siRNA-lipoplex technology from Silence Therapeutics (London, United Kingdom), STEMFECT™ from STEMGENT® (Cambridge, Mass.), and polyethylenimine (PEI) or protamine-based targeted and non-targeted delivery of nucleic acids (Aleku et al. Cancer Res. 2008 68:9788-9798; Strumberg et al. Int J Clin Pharmacol Ther 2012 50:76-78; Santel et al., Gene Ther 2006 13:1222-1234; Santel et al., Gene Ther 2006 13:1360-1370; Gutbier et al., Pulm Pharmacol. Ther. 2010 23:334-344; Kaufmann et al. Microvasc Res 2010 80:286-293 Weide et al. J Immunother. 2009 32:498-507; Weide et al. J Immunother. 2008 31:180-188; Pascolo Expert Opin. Biol. Ther. 4:1285-1294; Fotin-Mleczek et al., 2011 J. Immunother. 34:1-15; Song et al., Nature Biotechnol. 2005, 23:709-717; Peer et al., Proc Natl Acad Sci USA. 2007 6; 104:4095-4100; deFougerolles Hum Gene Ther. 2008 19:125-132, the contents of each of which are incorporated herein by reference in their entirety).

In some embodiments, such formulations may also be constructed or compositions altered such that they passively or actively are directed to different cell types in vivo, including but not limited to hepatocytes, immune cells, tumor cells, endothelial cells, antigen presenting cells, and leukocytes (Akinc et al. Mol Ther. 2010 18:1357-1364; Song et al., Nat Biotechnol. 2005 23:709-717; Judge et al., J Clin Invest. 2009 119:661-673; Kaufmann et al., Microvasc Res 2010 80:286-293; Santel et al., Gene Ther 2006 13:1222-1234; Santel et al., Gene Ther 2006 13:1360-1370; Gutbier et al., Pulm Pharmacol. Ther. 2010 23:334-344; Basha et al., Mol. Ther. 2011 19:2186-2200; Fenske and Cullis, Expert Opin Drug Deliv. 2008 5:25-44; Peer et al., Science. 2008 319:627-630; Peer and Lieberman, Gene Ther. 2011 18:1127-1133, the contents of each of which are incorporated herein by reference in their entirety). One example of passive targeting of formulations to liver cells includes the DLin-DMA, DLin-KC2-DMA and DLin-MC3-DMA-based lipid nanoparticle formulations, which have been shown to bind to apolipoprotein E and promote binding and uptake of these formulations into hepatocytes in vivo (Akinc et al. Mol Ther. 2010 18:1357-1364, the contents of which are incorporated herein by reference in their entirety). Formulations can also be selectively targeted through expression of different ligands on their surface as exemplified by, but not limited by, folate, transferrin, N-acetylgalactosamine (GalNAc), and antibody targeted approaches (Kolhatkar et al., Curr Drug Discov Technol. 2011 8:197-206; Musacchio and Torchilin, Front Biosci. 2011 16:1388-1412; Yu et al., Mol Membr Biol. 2010 27:286-298; Patil et al., Crit Rev Ther Drug Carrier Syst. 2008 25:1-61; Benoit et al., Biomacromolecules. 2011 12:2708-2714; Zhao et al., Expert Opin Drug Deliv. 2008 5:309-319; Akinc et al., Mol Ther. 2010 18:1357-1364; Srinivasan et al., Methods Mol Biol. 2012 820:105-116; Ben-Arie et al., Methods Mol Biol. 2012 757:497-507; Peer 2010 J Control Release. 20:63-68; Peer et al., Proc Natl Acad Sci USA. 2007 104:4095-4100; Kim et al., Methods Mol Biol. 2011 721:339-353; Subramanya et al., Mol Ther. 2010 18:2028-2037; Song et al., Nat Biotechnol. 2005 23:709-717; Peer et al., Science. 2008 319:627-630; Peer and Lieberman, Gene Ther. 2011 18:1127-1133, the contents of each of which are incorporated herein by reference in their entirety).

In some embodiments, the RNA (e.g., mRNA) vaccine is formulated as a solid lipid nanoparticle. A solid lipid nanoparticle (SLN) may be spherical with an average diameter between 10 to 1000 nm. SLNs possess a solid lipid core matrix that can solubilize lipophilic molecules and may be stabilized with surfactants and/or emulsifiers. In some embodiments, the lipid nanoparticle may be a self-assembly lipid-polymer nanoparticle (see Zhang et al., ACS Nano, 2008, 2, pp 1696-1702; the contents of which are herein incorporated by reference in their entirety). As a non-limiting example, the SLN may be the SLN described in International Patent Publication No. WO2013105101, the contents of which are herein incorporated by reference in their entirety. As another non-limiting example, the SLN may be made by the methods or processes described in International Patent Publication No. WO2013105101, the contents of which are herein incorporated by reference in their entirety.

Liposomes, lipoplexes, or lipid nanoparticles may be used to improve the efficacy of polynucleotides directed protein production as these formulations may be able to increase cell transfection by the RNA (e.g., mRNA) vaccine; and/or increase the translation of encoded protein. One such example involves the use of lipid encapsulation to enable the effective systemic delivery of polyplex plasmid DNA (Heyes et al., Mol Ther. 2007 15:713-720; the contents of which are incorporated herein by reference in their entirety). The liposomes, lipoplexes, or lipid nanoparticles may also be used to increase the stability of the polynucleotide.

In some embodiments, the RNA (e.g., mRNA) vaccines of the present disclosure can be formulated for controlled release and/or targeted delivery. As used herein, “controlled release” refers to a pharmaceutical composition or compound release profile that conforms to a particular pattern of release to effect a therapeutic outcome. In some embodiments, the RNA (e.g., mRNA) vaccines may be encapsulated into a delivery agent described herein and/or known in the art for controlled release and/or targeted delivery. As used herein, the term “encapsulate” means to enclose, surround or encase. As it relates to the formulation of the compounds of the disclosure, encapsulation may be substantial, complete or partial. The term “substantially encapsulated” means that at least greater than 50, 60, 70, 80, 85, 90, 95, 96, 97, 98, 99, 99.9, 99.99 or greater than 99.999% of the pharmaceutical composition or compound of the disclosure may be enclosed, surrounded or encased within the delivery agent. “Partially encapsulation” means that less than 10, 10, 20, 30, 40, 50% or less of the pharmaceutical composition or compound of the disclosure may be enclosed, surrounded or encased within the delivery agent. Advantageously, encapsulation may be determined by measuring the escape or the activity of the pharmaceutical composition or compound of the disclosure using fluorescence and/or electron micrograph. For example, at least 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 85, 90, 95, 96, 97, 98, 99, 99.9, 99.99 or greater than 99.999% of the pharmaceutical composition or compound of the disclosure are encapsulated in the delivery agent.

In some embodiments, the controlled release formulation may include, but is not limited to, tri-block co-polymers. As a non-limiting example, the formulation may include two different types of tri-block co-polymers (International Pub. No. WO2012131104 and WO2012131106, the contents of each of which are incorporated herein by reference in their entirety).

In some embodiments, the RNA (e.g., mRNA) vaccines may be encapsulated into a lipid nanoparticle or a rapidly eliminated lipid nanoparticle and the lipid nanoparticles or a rapidly eliminated lipid nanoparticle may then be encapsulated into a polymer, hydrogel and/or surgical sealant described herein and/or known in the art. As a non-limiting example, the polymer, hydrogel or surgical sealant may be PLGA, ethylene vinyl acetate (EVAc), poloxamer, GELSITE® (Nanotherapeutics, Inc. Alachua, Fla.), HYLENEX® (Halozyme Therapeutics, San Diego Calif.), surgical sealants such as fibrinogen polymers (Ethicon Inc. Cornelia, Ga.), TISSELL® (Baxter International, Inc Deerfield, Ill.), PEG-based sealants, and COSEAL® (Baxter International, Inc Deerfield, Ill.).

In some embodiments, the lipid nanoparticle may be encapsulated into any polymer known in the art which may form a gel when injected into a subject. As another non-limiting example, the lipid nanoparticle may be encapsulated into a polymer matrix which may be biodegradable.

In some embodiments, the RNA (e.g., mRNA) vaccine formulation for controlled release and/or targeted delivery may also include at least one controlled release coating. Controlled release coatings include, but are not limited to, OPADRY®, polyvinylpyrrolidone/vinyl acetate copolymer, polyvinylpyrrolidone, hydroxypropyl methylcellulose, hydroxypropyl cellulose, hydroxyethyl cellulose, EUDRAGIT RL®, EUDRAGIT RS® and cellulose derivatives such as ethylcellulose aqueous dispersions (AQUACOAT® and SURELEASE®).

In some embodiments, the RNA (e.g., mRNA) vaccine controlled release and/or targeted delivery formulation may comprise at least one degradable polyester which may contain polycationic side chains. Degradeable polyesters include, but are not limited to, poly(serine ester), poly(L-lactide-co-L-lysine), poly(4-hydroxy-L-proline ester), and combinations thereof. In some embodiments, the degradable polyesters may include a PEG conjugation to form a PEGylated polymer.

In some embodiments, the RNA (e.g., mRNA) vaccine controlled release and/or targeted delivery formulation comprising at least one polynucleotide may comprise at least one PEG and/or PEG related polymer derivatives as described in U.S. Pat. No. 8,404,222, the contents of which are incorporated herein by reference in their entirety.

In some embodiments, the RNA (e.g., mRNA) vaccine controlled release delivery formulation comprising at least one polynucleotide may be the controlled release polymer system described in US20130130348, the contents of which are incorporated herein by reference in their entirety.

In some embodiments, the RNA (e.g., mRNA) vaccines of the present disclosure may be encapsulated in a therapeutic nanoparticle, referred to herein as “therapeutic nanoparticle RNA (e.g., mRNA) vaccines.” Therapeutic nanoparticles may be formulated by methods described herein and known in the art such as, but not limited to, International Pub Nos. WO2010005740, WO2010030763, WO2010005721, WO2010005723, WO2012054923, U.S. Publication Nos. US20110262491, US20100104645, US20100087337, US20100068285, US20110274759, US20100068286, US20120288541, US20130123351 and US20130230567 and U.S. Pat. Nos. 8,206,747, 8,293,276, 8,318,208 and 8,318,211; the contents of each of which are herein incorporated by reference in their entirety. In some embodiments, therapeutic polymer nanoparticles may be identified by the methods described in US Pub No. US20120140790, the contents of which are herein incorporated by reference in their entirety.

In some embodiments, the therapeutic nanoparticle RNA (e.g., mRNA) vaccine may be formulated for sustained release. As used herein, “sustained release” refers to a pharmaceutical composition or compound that conforms to a release rate over a specific period of time. The period of time may include, but is not limited to, hours, days, weeks, months and years. As a non-limiting example, the sustained release nanoparticle may comprise a polymer and a therapeutic agent such as, but not limited to, the polynucleotides of the present disclosure (see International Pub No. 2010075072 and US Pub No. US20100216804, US20110217377 and US20120201859, the contents of each of which are incorporated herein by reference in their entirety). In another non-limiting example, the sustained release formulation may comprise agents which permit persistent bioavailability such as, but not limited to, crystals, macromolecular gels and/or particulate suspensions (see U.S. Patent Publication No. US20130150295, the contents of each of which are incorporated herein by reference in their entirety).

In some embodiments, the therapeutic nanoparticle RNA (e.g., mRNA) vaccines may be formulated to be target specific. As a non-limiting example, the therapeutic nanoparticles may include a corticosteroid (see International Pub. No. WO2011084518, the contents of which are incorporated herein by reference in their entirety). As a non-limiting example, the therapeutic nanoparticles may be formulated in nanoparticles described in International Pub No. WO2008121949, WO2010005726, WO2010005725, WO2011084521 and US Pub No. US20100069426, US20120004293 and US20100104655, the contents of each of which are incorporated herein by reference in their entirety.

In some embodiments, the nanoparticles of the present disclosure may comprise a polymeric matrix. As a non-limiting example, the nanoparticle may comprise two or more polymers such as, but not limited to, polyethylenes, polycarbonates, polyanhydrides, polyhydroxyacids, polypropylfumerates, polycaprolactones, polyamides, polyacetals, polyethers, polyesters, poly(orthoesters), polycyanoacrylates, polyvinyl alcohols, polyurethanes, polyphosphazenes, polyacrylates, polymethacrylates, polycyanoacrylates, polyureas, polystyrenes, polyamines, polylysine, poly(ethylene imine), poly(serine ester), poly(L-lactide-co-L-lysine), poly(4-hydroxy-L-proline ester) or combinations thereof.

In some embodiments, the therapeutic nanoparticle comprises a diblock copolymer. In some embodiments, the diblock copolymer may include PEG in combination with a polymer such as, but not limited to, polyethylenes, polycarbonates, polyanhydrides, polyhydroxyacids, polypropylfumerates, polycaprolactones, polyamides, polyacetals, polyethers, polyesters, poly(orthoesters), polycyanoacrylates, polyvinyl alcohols, polyurethanes, polyphosphazenes, polyacrylates, polymethacrylates, polycyanoacrylates, polyureas, polystyrenes, polyamines, polylysine, poly(ethylene imine), poly(serine ester), poly(L-lactide-co-L-lysine), poly(4-hydroxy-L-proline ester) or combinations thereof. In yet another embodiment, the diblock copolymer may be a high-X diblock copolymer such as those described in International Patent Publication No. WO2013120052, the contents of which are incorporated herein by reference in their entirety.

As a non-limiting example the therapeutic nanoparticle comprises a PLGA-PEG block copolymer (see U.S. Publication No. US20120004293 and U.S. Pat. No. 8,236,330, each of which is herein incorporated by reference in their entirety). In another non-limiting example, the therapeutic nanoparticle is a stealth nanoparticle comprising a diblock copolymer of PEG and PLA or PEG and PLGA (see U.S. Pat. No. 8,246,968 and International Publication No. WO2012166923, the contents of each of which are herein incorporated by reference in their entirety). In yet another non-limiting example, the therapeutic nanoparticle is a stealth nanoparticle or a target-specific stealth nanoparticle as described in U.S. Patent Publication No. US20130172406, the contents of which are herein incorporated by reference in their entirety.

In some embodiments, the therapeutic nanoparticle may comprise a multiblock copolymer (see e.g., U.S. Pat. Nos. 8,263,665 and 8,287,910 and U.S. Patent Pub. No. US20130195987, the contents of each of which are herein incorporated by reference in their entirety).

In yet another non-limiting example, the lipid nanoparticle comprises the block copolymer PEG-PLGA-PEG (see e.g., the thermosensitive hydrogel (PEG-PLGA-PEG) was used as a TGF-beta1 gene delivery vehicle in Lee et al. Thermosensitive Hydrogel as a TGF-β1 Gene Delivery Vehicle Enhances Diabetic Wound Healing. Pharmaceutical Research, 2003 20(12): 1995-2000; as a controlled gene delivery system in Li et al. Controlled Gene Delivery System Based on Thermosensitive Biodegradable Hydrogel. Pharmaceutical Research 2003 20:884-888; and Chang et al., Non-ionic amphiphilic biodegradable PEG-PLGA-PEG copolymer enhances gene delivery efficiency in rat skeletal muscle. J Controlled Release. 2007 118:245-253, the contents of each of which are herein incorporated by reference in their entirety). The RNA (e.g., mRNA) vaccines of the present disclosure may be formulated in lipid nanoparticles comprising the PEG-PLGA-PEG block copolymer.

In some embodiments, the therapeutic nanoparticle may comprise a multiblock copolymer (see e.g., U.S. Pat. Nos. 8,263,665 and 8,287,910 and U.S. Patent Pub. No. US20130195987, the contents of each of which are herein incorporated by reference in their entirety).

In some embodiments, the block copolymers described herein may be included in a polyion complex comprising a non-polymeric micelle and the block copolymer. (see e.g., U.S. Publication No. 20120076836, the contents of which are herein incorporated by reference in their entirety).

In some embodiments, the therapeutic nanoparticle may comprise at least one acrylic polymer. Acrylic polymers include but are not limited to, acrylic acid, methacrylic acid, acrylic acid and methacrylic acid copolymers, methyl methacrylate copolymers, ethoxyethyl methacrylates, cyanoethyl methacrylate, amino alkyl methacrylate copolymer, poly(acrylic acid), poly(methacrylic acid), polycyanoacrylates and combinations thereof.

In some embodiments, the therapeutic nanoparticles may comprise at least one poly(vinyl ester) polymer. The poly(vinyl ester) polymer may be a copolymer such as a random copolymer. As a non-limiting example, the random copolymer may have a structure such as those described in International Application No. WO2013032829 or U.S. Patent Publication No US20130121954, the contents of each of which are herein incorporated by reference in their entirety. In some embodiments, the poly(vinyl ester) polymers may be conjugated to the polynucleotides described herein.

In some embodiments, the therapeutic nanoparticle may comprise at least one diblock copolymer. The diblock copolymer may be, but it not limited to, a poly(lactic) acid-poly(ethylene)glycol copolymer (see, e.g., International Patent Publication No. WO2013044219, the contents of which are herein incorporated by reference in their entirety). As a non-limiting example, the therapeutic nanoparticle may be used to treat cancer (see International publication No. WO2013044219, the contents of which are herein incorporated by reference in their entirety).

In some embodiments, the therapeutic nanoparticles may comprise at least one cationic polymer described herein and/or known in the art.

In some embodiments, the therapeutic nanoparticles may comprise at least one amine-containing polymer such as, but not limited to polylysine, polyethylene imine, poly(amidoamine) dendrimers, poly(beta-amino esters) (see, e.g., U.S. Pat. No. 8,287,849, the contents of which are herein incorporated by reference in their entirety) and combinations thereof.

In some embodiments, the nanoparticles described herein may comprise an amine cationic lipid such as those described in International Patent Application No. WO2013059496, the contents of which are herein incorporated by reference in their entirety. In some embodiments, the cationic lipids may have an amino-amine or an amino-amide moiety.

In some embodiments, the therapeutic nanoparticles may comprise at least one degradable polyester which may contain polycationic side chains. Degradeable polyesters include, but are not limited to, poly(serine ester), poly(L-lactide-co-L-lysine), poly(4-hydroxy-L-proline ester), and combinations thereof. In some embodiments, the degradable polyesters may include a PEG conjugation to form a PEGylated polymer.

In some embodiments, the synthetic nanocarriers may contain an immunostimulatory agent to enhance the immune response from delivery of the synthetic nanocarrier. As a non-limiting example, the synthetic nanocarrier may comprise a Th1 immunostimulatory agent, which may enhance a Th1-based response of the immune system (see International Pub No. WO2010123569 and U.S. Publication No. US20110223201, the contents of each of which are herein incorporated by reference in their entirety).

In some embodiments, the synthetic nanocarriers may be formulated for targeted release. In some embodiments, the synthetic nanocarrier is formulated to release the polynucleotides at a specified pH and/or after a desired time interval. As a non-limiting example, the synthetic nanoparticle may be formulated to release the RNA (e.g., mRNA) vaccines after 24 hours and/or at a pH of 4.5 (see International Publication Nos. WO2010138193 and WO2010138194 and US Pub Nos. US20110020388 and US20110027217, each of which is herein incorporated by reference in their entireties).

In some embodiments, the synthetic nanocarriers may be formulated for controlled and/or sustained release of the polynucleotides described herein. As a non-limiting example, the synthetic nanocarriers for sustained release may be formulated by methods known in the art, described herein and/or as described in International Pub No. WO2010138192 and US Pub No. 20100303850, each of which is herein incorporated by reference in their entirety.

In some embodiments, the RNA (e.g., mRNA) vaccine may be formulated for controlled and/or sustained release wherein the formulation comprises at least one polymer that is a crystalline side chain (CYSC) polymer. CYSC polymers are described in U.S. Pat. No. 8,399,007, herein incorporated by reference in its entirety.

In some embodiments, the synthetic nanocarrier may be formulated for use as a vaccine. In some embodiments, the synthetic nanocarrier may encapsulate at least one polynucleotide which encode at least one antigen. As a non-limiting example, the synthetic nanocarrier may include at least one antigen and an excipient for a vaccine dosage form (see International Publication No. WO2011150264 and U.S. Publication No. US20110293723, the contents of each of which are herein incorporated by reference in their entirety). As another non-limiting example, a vaccine dosage form may include at least two synthetic nanocarriers with the same or different antigens and an excipient (see International Publication No. WO2011150249 and U.S. Publication No. US20110293701, the contents of each of which are herein incorporated by reference in their entirety). The vaccine dosage form may be selected by methods described herein, known in the art and/or described in International Publication No. WO2011150258 and U.S. Publication No. US20120027806, the contents of each of which are herein incorporated by reference in their entirety).

In some embodiments, the synthetic nanocarrier may comprise at least one polynucleotide which encodes at least one adjuvant. As non-limiting example, the adjuvant may comprise dimethyldioctadecylammonium-bromide, dimethyldioctadecylammonium-chloride, dimethyldioctadecylammonium-phosphate or dimethyldioctadecylammonium-acetate (DDA) and an apolar fraction or part of said apolar fraction of a total lipid extract of a mycobacterium (see, e.g., U.S. Pat. No. 8,241,610, the content of which is herein incorporated by reference in its entirety). In some embodiments, the synthetic nanocarrier may comprise at least one polynucleotide and an adjuvant. As a non-limiting example, the synthetic nanocarrier comprising and adjuvant may be formulated by the methods described in International Publication No. WO2011150240 and U.S. Publication No. US20110293700, the contents of each of which are herein incorporated by reference in their entirety.

In some embodiments, the synthetic nanocarrier may encapsulate at least one polynucleotide that encodes a peptide, fragment or region from a virus. As a non-limiting example, the synthetic nanocarrier may include, but is not limited to, any of the nanocarriers described in International Publication No. WO2012024621, WO201202629, WO2012024632 and U.S. Publication No. US20120064110, US20120058153 and US20120058154, the contents of each of which are herein incorporated by reference in their entirety.

In some embodiments, the synthetic nanocarrier may be coupled to a polynucleotide which may be able to trigger a humoral and/or cytotoxic T lymphocyte (CTL) response (see, e.g., International Publication No. WO2013019669, the contents of which are herein incorporated by reference in their entirety).

In some embodiments, the RNA (e.g., mRNA) vaccine may be encapsulated in, linked to and/or associated with zwitterionic lipids. Non-limiting examples of zwitterionic lipids and methods of using zwitterionic lipids are described in U.S. Patent Publication No. US20130216607, the contents of which are herein incorporated by reference in their entirety. In some aspects, the zwitterionic lipids may be used in the liposomes and lipid nanoparticles described herein.

In some embodiments, the RNA (e.g., mRNA) vaccine may be formulated in colloid nanocarriers as described in U.S. Patent Publication No. US20130197100, the contents of which are herein incorporated by reference in their entirety.

In some embodiments, the nanoparticle may be optimized for oral administration. The nanoparticle may comprise at least one cationic biopolymer such as, but not limited to, chitosan or a derivative thereof. As a non-limiting example, the nanoparticle may be formulated by the methods described in U.S. Publication No. 20120282343, the contents of which are herein incorporated by reference in their entirety.

In some embodiments, LNPs comprise the lipid KL52 (an amino-lipid disclosed in U.S. Application Publication No. 2012/0295832, the contents of which are herein incorporated by reference in their entirety. Activity and/or safety (as measured by examining one or more of ALT/AST, white blood cell count and cytokine induction, for example) of LNP administration may be improved by incorporation of such lipids. LNPs comprising KL52 may be administered intravenously and/or in one or more doses. In some embodiments, administration of LNPs comprising KL52 results in equal or improved mRNA and/or protein expression as compared to LNPs comprising MC3.

In some embodiments, RNA (e.g., mRNA) vaccine may be delivered using smaller LNPs. Such particles may comprise a diameter from below 0.1 um up to 100 nm such as, but not limited to, less than 0.1 um, less than 1.0 um, less than 5 um, less than 10 um, less than 15 um, less than 20 um, less than 25 um, less than 30 um, less than 35 um, less than 40 um, less than 50 um, less than 55 um, less than 60 um, less than 65 um, less than 70 um, less than 75 um, less than 80 um, less than 85 um, less than 90 um, less than 95 um, less than 100 um, less than 125 um, less than 150 um, less than 175 um, less than 200 um, less than 225 um, less than 250 um, less than 275 um, less than 300 um, less than 325 um, less than 350 um, less than 375 um, less than 400 um, less than 425 um, less than 450 um, less than 475 um, less than 500 um, less than 525 um, less than 550 um, less than 575 um, less than 600 um, less than 625 um, less than 650 um, less than 675 um, less than 700 um, less than 725 um, less than 750 um, less than 775 um, less than 800 um, less than 825 um, less than 850 um, less than 875 um, less than 900 um, less than 925 um, less than 950 um, less than 975 um, or less than 1000 um.

In some embodiments, RNA (e.g., mRNA) vaccines may be delivered using smaller LNPs, which may comprise a diameter from about 1 nm to about 100 nm, from about 1 nm to about 10 nm, about 1 nm to about 20 nm, from about 1 nm to about 30 nm, from about 1 nm to about 40 nm, from about 1 nm to about 50 nm, from about 1 nm to about 60 nm, from about 1 nm to about 70 nm, from about 1 nm to about 80 nm, from about 1 nm to about 90 nm, from about 5 nm to about from 100 nm, from about 5 nm to about 10 nm, about 5 nm to about 20 nm, from about 5 nm to about 30 nm, from about 5 nm to about 40 nm, from about 5 nm to about 50 nm, from about 5 nm to about 60 nm, from about 5 nm to about 70 nm, from about 5 nm to about 80 nm, from about 5 nm to about 90 nm, about 10 to about 50 nm, from about 20 to about 50 nm, from about 30 to about 50 nm, from about 40 to about 50 nm, from about 20 to about 60 nm, from about 30 to about 60 nm, from about 40 to about 60 nm, from about 20 to about 70 nm, from about 30 to about 70 nm, from about 40 to about 70 nm, from about 50 to about 70 nm, from about 60 to about 70 nm, from about 20 to about 80 nm, from about 30 to about 80 nm, from about 40 to about 80 nm, from about 50 to about 80 nm, from about 60 to about 80 nm, from about 20 to about 90 nm, from about 30 to about 90 nm, from about 40 to about 90 nm, from about 50 to about 90 nm, from about 60 to about 90 nm and/or from about 70 to about 90 nm.

In some embodiments, such LNPs are synthesized using methods comprising microfluidic mixers. Examples of microfluidic mixers may include, but are not limited to, a slit interdigital micromixer including, but not limited to those manufactured by Microinnova (Allerheiligen bei Wildon, Austria) and/or a staggered herringbone micromixer (SHM) (Zhigaltsev, I. V. et al., Bottom-up design and synthesis of limit size lipid nanoparticle systems with aqueous and triglyceride cores using millisecond microfluidic mixing have been published (Langmuir. 2012. 28:3633-40; Belliveau, N. M. et al., Microfluidic synthesis of highly potent limit-size lipid nanoparticles for in vivo delivery of siRNA. Molecular Therapy-Nucleic Acids. 2012. 1:e37; Chen, D. et al., Rapid discovery of potent siRNA-containing lipid nanoparticles enabled by controlled microfluidic formulation. J Am Chem Soc. 2012. 134(16):6948-51, the contents of each of which are herein incorporated by reference in their entirety). In some embodiments, methods of LNP generation comprising SHM, further comprise the mixing of at least two input streams wherein mixing occurs by microstructure-induced chaotic advection (MICA). According to this method, fluid streams flow through channels present in a herringbone pattern, causing rotational flow and folding the fluids around each other. This method may also comprise a surface for fluid mixing wherein the surface changes orientations during fluid cycling. Methods of generating LNPs using SHM include those disclosed in U.S. Application Publication Nos. 2004/0262223 and 2012/0276209, the contents of each of which are herein incorporated by reference in their entirety.

In some embodiments, the RNA (e.g., mRNA) vaccine of the present disclosure may be formulated in lipid nanoparticles created using a micromixer such as, but not limited to, a Slit Interdigital Microstructured Mixer (SIMM-V2) or a Standard Slit Interdigital Micro Mixer (SSIMM) or Caterpillar (CPMM) or Impinging-jet (IJMM) from the Institut für Mikrotechnik Mainz GmbH, Mainz Germany.

In some embodiments, the RNA (e.g., mRNA) vaccines of the present disclosure may be formulated in lipid nanoparticles created using microfluidic technology (see, e.g., Whitesides, George M. The Origins and the Future of Microfluidics. Nature, 2006 442: 368-373; and Abraham et al. Chaotic Mixer for Microchannels. Science, 2002 295: 647-651; each of which is herein incorporated by reference in its entirety). As a non-limiting example, controlled microfluidic formulation includes a passive method for mixing streams of steady pressure-driven flows in micro channels at a low Reynolds number (see, e.g., Abraham et al. Chaotic Mixer for Microchannels. Science, 2002 295: 647-651, the contents of which are herein incorporated by reference in their entirety).

In some embodiments, the RNA (e.g., mRNA) vaccines of the present disclosure may be formulated in lipid nanoparticles created using a micromixer chip such as, but not limited to, those from Harvard Apparatus (Holliston, Mass.) or Dolomite Microfluidics (Royston, UK). A micromixer chip can be used for rapid mixing of two or more fluid streams with a split and recombine mechanism.

In some embodiments, the RNA (e.g., mRNA) vaccines of the disclosure may be formulated for delivery using the drug encapsulating microspheres described in International Patent Publication No. WO2013063468 or U.S. Pat. No. 8,440,614, the contents of each of which are herein incorporated by reference in their entirety. The microspheres may comprise a compound of the formula (I), (II), (III), (IV), (V) or (VI) as described in International Patent Publication No. WO2013063468, the contents of which are herein incorporated by reference in their entirety. In some embodiments, the amino acid, peptide, polypeptide, and/or lipids are useful in delivering the RNA (e.g., mRNA) vaccines of the disclosure to cells (see International Patent Publication No. WO2013063468, the contents of which are herein incorporated by reference in their entirety).

In some embodiments, the RNA (e.g., mRNA) vaccines of the disclosure may be formulated in lipid nanoparticles having a diameter from about 10 to about 100 nm such as, but not limited to, about 10 to about 20 nm, about 10 to about 30 nm, about 10 to about 40 nm, about 10 to about 50 nm, about 10 to about 60 nm, about 10 to about 70 nm, about 10 to about 80 nm, about 10 to about 90 nm, about 20 to about 30 nm, about 20 to about 40 nm, about 20 to about 50 nm, about 20 to about 60 nm, about 20 to about 70 nm, about 20 to about 80 nm, about 20 to about 90 nm, about 20 to about 100 nm, about 30 to about 40 nm, about 30 to about 50 nm, about 30 to about 60 nm, about 30 to about 70 nm, about 30 to about 80 nm, about 30 to about 90 nm, about 30 to about 100 nm, about 40 to about 50 nm, about 40 to about 60 nm, about 40 to about 70 nm, about 40 to about 80 nm, about 40 to about 90 nm, about 40 to about 100 nm, about 50 to about 60 nm, about 50 to about 70 nm about 50 to about 80 nm, about 50 to about 90 nm, about 50 to about 100 nm, about 60 to about 70 nm, about 60 to about 80 nm, about 60 to about 90 nm, about 60 to about 100 nm, about 70 to about 80 nm, about 70 to about 90 nm, about 70 to about 100 nm, about 80 to about 90 nm, about 80 to about 100 nm and/or about 90 to about 100 nm.

In some embodiments, the lipid nanoparticles may have a diameter from about 10 to 500 nm.

In some embodiments, the lipid nanoparticle may have a diameter greater than 100 nm, greater than 150 nm, greater than 200 nm, greater than 250 nm, greater than 300 nm, greater than 350 nm, greater than 400 nm, greater than 450 nm, greater than 500 nm, greater than 550 nm, greater than 600 nm, greater than 650 nm, greater than 700 nm, greater than 750 nm, greater than 800 nm, greater than 850 nm, greater than 900 nm, greater than 950 nm or greater than 1000 nm.

In some embodiments, the lipid nanoparticle may be a limit size lipid nanoparticle described in International Patent Publication No. WO2013059922, the contents of which are herein incorporated by reference in their entirety. The limit size lipid nanoparticle may comprise a lipid bilayer surrounding an aqueous core or a hydrophobic core; where the lipid bilayer may comprise a phospholipid such as, but not limited to, diacylphosphatidylcholine, a diacylphosphatidylethanolamine, a ceramide, a sphingomyelin, a dihydrosphingomyelin, a cephalin, a cerebroside, a C8-C20 fatty acid diacylphophatidylcholine, and 1-palmitoyl-2-oleoyl phosphatidylcholine (POPC). In some embodiments, the limit size lipid nanoparticle may comprise a polyethylene glycol-lipid such as, but not limited to, DLPE-PEG, DMPE-PEG, DPPC-PEG and DSPE-PEG.

In some embodiments, the RNA (e.g., mRNA) vaccines may be delivered, localized and/or concentrated in a specific location using the delivery methods described in International Patent Publication No. WO2013063530, the contents of which are herein incorporated by reference in their entirety. As a non-limiting example, a subject may be administered an empty polymeric particle prior to, simultaneously with or after delivering the RNA (e.g., mRNA) vaccines to the subject. The empty polymeric particle undergoes a change in volume once in contact with the subject and becomes lodged, embedded, immobilized or entrapped at a specific location in the subject.

In some embodiments, the RNA (e.g., mRNA) vaccines may be formulated in an active substance release system (see, e.g., U.S. Patent Publication No. US20130102545, the contents of which are herein incorporated by reference in their entirety). The active substance release system may comprise 1) at least one nanoparticle bonded to an oligonucleotide inhibitor strand which is hybridized with a catalytically active nucleic acid and 2) a compound bonded to at least one substrate molecule bonded to a therapeutically active substance (e.g., polynucleotides described herein), where the therapeutically active substance is released by the cleavage of the substrate molecule by the catalytically active nucleic acid.

In some embodiments, the RNA (e.g., mRNA) vaccines may be formulated in a nanoparticle comprising an inner core comprising a non-cellular material and an outer surface comprising a cellular membrane. The cellular membrane may be derived from a cell or a membrane derived from a virus. As a non-limiting example, the nanoparticle may be made by the methods described in International Patent Publication No. WO2013052167, the contents of which are herein incorporated by reference in their entirety. As another non-limiting example, the nanoparticle described in International Patent Publication No. WO2013052167, the contents of which are herein incorporated by reference in their entirety, may be used to deliver the RNA (e.g., mRNA) vaccines described herein.

In some embodiments, the RNA (e.g., mRNA) vaccines may be formulated in porous nanoparticle-supported lipid bilayers (protocells). Protocells are described in International Patent Publication No. WO2013056132, the contents of which are herein incorporated by reference in their entirety.

In some embodiments, the RNA (e.g., mRNA) vaccines described herein may be formulated in polymeric nanoparticles as described in or made by the methods described in U.S. Pat. Nos. 8,420,123 and 8,518,963 and European Patent No. EP2073848B1, the contents of each of which are herein incorporated by reference in their entirety. As a non-limiting example, the polymeric nanoparticle may have a high glass transition temperature such as the nanoparticles described in or nanoparticles made by the methods described in U.S. Pat. No. 8,518,963, the contents of which are herein incorporated by reference in their entirety. As another non-limiting example, the polymer nanoparticle for oral and parenteral formulations may be made by the methods described in European Patent No. EP2073848B1, the contents of which are herein incorporated by reference in their entirety.

In some embodiments, the RNA (e.g., mRNA) vaccines described herein may be formulated in nanoparticles used in imaging. The nanoparticles may be liposome nanoparticles such as those described in U.S. Patent Publication No US20130129636, herein incorporated by reference in its entirety. As a non-limiting example, the liposome may comprise gadolinium(III)2-{4,7-bis-carboxymethyl-10-[(N,N-distearylamidomethyl-N′-amido-methyl]-1,4,7,10-tetra-azacyclododec-1-yl}-acetic acid and a neutral, fully saturated phospholipid component (see, e.g., U.S. Patent Publication No US20130129636, the contents of which are herein incorporated by reference in their entirety).

In some embodiments, the nanoparticles which may be used in the present disclosure are formed by the methods described in U.S. Patent Application No. US20130130348, the contents of which are herein incorporated by reference in their entirety.

The nanoparticles of the present disclosure may further include nutrients such as, but not limited to, those which deficiencies can lead to health hazards from anemia to neural tube defects (see, e.g., the nanoparticles described in International Patent Publication No WO2013072929, the contents of which are herein incorporated by reference in their entirety). As a non-limiting example, the nutrient may be iron in the form of ferrous, ferric salts or elemental iron, iodine, folic acid, vitamins or micronutrients.

In some embodiments, the RNA (e.g., mRNA) vaccines of the present disclosure may be formulated in a swellable nanoparticle. The swellable nanoparticle may be, but is not limited to, those described in U.S. Pat. No. 8,440,231, the contents of which are herein incorporated by reference in their entirety. As a non-limiting embodiment, the swellable nanoparticle may be used for delivery of the RNA (e.g., mRNA) vaccines of the present disclosure to the pulmonary system (see, e.g., U.S. Pat. No. 8,440,231, the contents of which are herein incorporated by reference in their entirety).

The RNA (e.g., mRNA) vaccines of the present disclosure may be formulated in polyanhydride nanoparticles such as, but not limited to, those described in U.S. Pat. No. 8,449,916, the contents of which are herein incorporated by reference in their entirety.

The nanoparticles and microparticles of the present disclosure may be geometrically engineered to modulate macrophage and/or the immune response. In some embodiments, the geometrically engineered particles may have varied shapes, sizes and/or surface charges in order to incorporated the polynucleotides of the present disclosure for targeted delivery such as, but not limited to, pulmonary delivery (see, e.g., International Publication No WO2013082111, the contents of which are herein incorporated by reference in their entirety). Other physical features the geometrically engineering particles may have include, but are not limited to, fenestrations, angled arms, asymmetry and surface roughness, and charge which can alter the interactions with cells and tissues. As a non-limiting example, nanoparticles of the present disclosure may be made by the methods described in International Publication No WO2013082111, the contents of which are herein incorporated by reference in their entirety.

In some embodiments, the nanoparticles of the present disclosure may be water soluble nanoparticles such as, but not limited to, those described in International Publication No. WO2013090601, the contents of which are herein incorporated by reference in their entirety. The nanoparticles may be inorganic nanoparticles which have a compact and zwitterionic ligand in order to exhibit good water solubility. The nanoparticles may also have small hydrodynamic diameters (HD), stability with respect to time, pH, and salinity and a low level of non-specific protein binding.

In some embodiments the nanoparticles of the present disclosure may be developed by the methods described in U.S. Patent Publication No. US20130172406, the contents of which are herein incorporated by reference in their entirety.

In some embodiments, the nanoparticles of the present disclosure are stealth nanoparticles or target-specific stealth nanoparticles such as, but not limited to, those described in U.S. Patent Publication No. US20130172406, the contents of which are herein incorporated by reference in their entirety. The nanoparticles of the present disclosure may be made by the methods described in U.S. Patent Publication No. US20130172406, the contents of which are herein incorporated by reference in their entirety.

In some embodiments, the stealth or target-specific stealth nanoparticles may comprise a polymeric matrix. The polymeric matrix may comprise two or more polymers such as, but not limited to, polyethylenes, polycarbonates, polyanhydrides, polyhydroxyacids, polypropylfumerates, polycaprolactones, polyamides, polyacetals, polyethers, polyesters, poly(orthoesters), polycyanoacrylates, polyvinyl alcohols, polyurethanes, polyphosphazenes, polyacrylates, polymethacrylates, polycyanoacrylates, polyureas, polystyrenes, polyamines, polyesters, polyanhydrides, polyethers, polyurethanes, polymethacrylates, polyacrylates, polycyanoacrylates or combinations thereof.

In some embodiments, the nanoparticle may be a nanoparticle-nucleic acid hybrid structure having a high density nucleic acid layer. As a non-limiting example, the nanoparticle-nucleic acid hybrid structure may made by the methods described in U.S. Patent Publication No. US20130171646, the contents of which are herein incorporated by reference in their entirety. The nanoparticle may comprise a nucleic acid such as, but not limited to, polynucleotides described herein and/or known in the art.

At least one of the nanoparticles of the present disclosure may be embedded in the core of a nanostructure or coated with a low density porous 3-D structure or coating which is capable of carrying or associating with at least one payload within or on the surface of the nanostructure. Non-limiting examples of the nanostructures comprising at least one nanoparticle are described in International Patent Publication No. WO2013123523, the contents of which are herein incorporated by reference in their entirety.

In some embodiments the RNA (e.g., mRNA) vaccine may be associated with a cationic or polycationic compounds, including protamine, nucleoline, spermine or spermidine, or other cationic peptides or proteins, such as poly-L-lysine (PLL), polyarginine, basic polypeptides, cell penetrating peptides (CPPs), including HIV-binding peptides, HIV-1 Tat (HIV), Tat-derived peptides, Penetratin, VP²² derived or analog peptides, Pestivirus Erns, HSINV, VP²² (Herpes simplex), MAP, KALA or protein transduction domains (PTDs), PpT620, prolin-rich peptides, arginine-rich peptides, lysine-rich peptides, MPG-peptide(s), Pep-1, L-oligomers, Calcitonin peptide(s), Antennapedia-derived peptides (particularly from Drosophila antennapedia), pAntp, plsl, FGF, Lactoferrin, Transportan, Buforin-2, Bac715-24, SynB, SynB, pVEC, hCT-derived peptides, SAP, histones, cationic polysaccharides, for example chitosan, polybrene, cationic polymers, e.g. polyethyleneimine (PEI), cationic lipids, e.g. DOTMA: [1-(2,3-sioleyloxy)propyl)]-N,N,N-trimethylammonium chloride, DMRIE, di-C14-amidine, DOTIM, SAINT, DC-Chol, BGTC, CTAP, DOPC, DODAP, DOPE: Dioleyl phosphatidylethanol-amine, DOSPA, DODAB, DOIC, DMEPC, DOGS: Dioctadecylamidoglicylspermin, DIMRI: Dimyristooxypropyl dimethyl hydroxyethyl ammonium bromide, DOTAP: dioleoyloxy-3-(trimethylammonio)propane, DC-6-14: O,O-ditetradecanoyl-N-.alpha.-trimethylammonioacetyl)diethanolamine chloride, CLIP 1: rac-[(2,3-dioctadecyloxypropyl)(2-hydroxyethyl)]-dimethylammonium chloride, CLIP6: rac-[2(2,3-dihexadecyloxypropyloxymethyloxy)ethyl]-trimethylammonium, CLIP9: rac-[2(2,3-dihexadecyloxypropyloxysuccinyloxy)ethyl]-trimethylammo-nium, oligofectamine, or cationic or polycationic polymers, e.g. modified polyaminoacids, such as beta-aminoacid-polymers or reversed polyamides, etc., modified polyethylenes, such as PVP (poly(N-ethyl-4-vinylpyridinium bromide)), etc., modified acrylates, such as pDMAEMA (poly(dimethylaminoethyl methylacrylate)), etc., modified amidoamines such as pAMAM (poly(amidoamine)), etc., modified polybetaminoester (PBAE), such as diamine end modified 1,4 butanediol diacrylate-co-5-amino-1-pentanol polymers, etc., dendrimers, such as polypropylamine dendrimers or pAMAM based dendrimers, etc., polyimine(s), such as PEI: poly(ethyleneimine), poly(propyleneimine), etc., polyallylamine, sugar backbone based polymers, such as cyclodextrin based polymers, dextran based polymers, chitosan, etc., silan backbone based polymers, such as PMOXA-PDMS copolymers, etc., blockpolymers consisting of a combination of one or more cationic blocks (e.g. selected from a cationic polymer as mentioned above) and of one or more hydrophilic or hydrophobic blocks (e.g. polyethyleneglycole), etc.

In other embodiments the RNA (e.g., mRNA) vaccine is not associated with a cationic or polycationic compounds.

In some embodiments, a nanoparticle comprises compounds of Formula (I):

or a salt or isomer thereof, wherein:

R₁ is selected from the group consisting of C₅₋₃₀ alkyl, C₅₋₂₀ alkenyl, —R*YR″, —YR″, and —R″M′R′;

R₂ and R₃ are independently selected from the group consisting of H, C₁₋₁₄ alkyl, C₂₋₁₄ alkenyl, —R*YR″, —YR″, and —R*OR″, or R₂ and R₃, together with the atom to which they are attached, form a heterocycle or carbocycle;

R₄ is selected from the group consisting of a C₃₋₆ carbocycle, —(CH₂)_(n)Q, —(CH₂)_(n)CHQR, —CHQR, —CQ(R)₂, and unsubstituted C₁₋₆ alkyl, where Q is selected from a carbocycle, heterocycle, —OR, —O(CH₂)_(n)N(R)₂, —C(O)OR, —OC(O)R, —CX₃, —CX₂H, —CXH₂, —CN, —N(R)₂, —C(O)N(R)₂, —N(R)C(O)R, —N(R)S(O)₂R, —N(R)C(O)N(R)₂, —N(R)C(S)N(R)₂, —N(R)R₈, —O(CH₂)_(n)OR, —N(R)C(═NR₉)N(R)₂, —N(R)C(═CHR₉)N(R)₂, —OC(O)N(R)₂, —N(R)C(O)OR, —N(OR)C(O)R, —N(OR)S(O)₂R, —N(OR)C(O)OR, —N(OR)C(O)N(R)₂, —N(OR)C(S)N(R)₂, —N(OR)C(═NR₉)N(R)₂, —N(OR)C(═CHR₉)N(R)₂, —C(═NR₉)N(R)₂, —C(═NR₉)R, —C(O)N(R)OR, and —C(R)N(R)₂C(O)OR, and each n is independently selected from 1, 2, 3, 4, and 5;

each R₅ is independently selected from the group consisting of C₁₋₃ alkyl, C₂₋₃ alkenyl, and H;

each R₆ is independently selected from the group consisting of C₁₋₃ alkyl, C₂₋₃ alkenyl, and H;

M and M′ are independently selected from —C(O)O—, —OC(O)—, —C(O)N(R′)—,

—N(R′)C(O)—, —C(O)—, —C(S)—, —C(S)S—, —SC(S)—, —CH(OH)—, —P(O)(OR′)O—, —S(O)₂—, —S—S—, an aryl group, and a heteroaryl group;

R₇ is selected from the group consisting of C₁₋₃ alkyl, C₂₋₃ alkenyl, and H;

R₈ is selected from the group consisting of C₃₋₆ carbocycle and heterocycle;

R₉ is selected from the group consisting of H, CN, NO₂, C₁₋₆ alkyl, —OR, —S(O)₂R, —S(O)₂N(R)₂, C₂₋₆ alkenyl, C₃₋₆ carbocycle and heterocycle;

each R is independently selected from the group consisting of C₁₋₃ alkyl, C₂₋₃ alkenyl, and H;

each R′ is independently selected from the group consisting of C₁₋₁₈ alkyl, C₂₋₁₈ alkenyl, —R*YR″, —YR″, and H;

-   -   each R″ is independently selected from the group consisting of         C₃₋₁₄ alkyl and C₃₋₁₄ alkenyl;     -   each R* is independently selected from the group consisting of         C₁₋₁₂ alkyl and C₂₋₁₂ alkenyl;     -   each Y is independently a C₃₋₆ carbocycle;     -   each X is independently selected from the group consisting of F,         Cl, Br, and I; and     -   m is selected from 5, 6, 7, 8, 9, 10, 11, 12, and 13.

In some embodiments, a subset of compounds of Formula (I) includes those in which when R₄ is —(CH₂)_(n)Q, —(CH₂)_(n)CHQR, —CHQR, or —CQ(R)₂, then (i) Q is not —N(R)₂ when n is 1, 2, 3, 4 or 5, or (ii) Q is not 5, 6, or 7-membered heterocycloalkyl when n is 1 or 2.

In some embodiments, another subset of compounds of Formula (I) includes those in which

R₁ is selected from the group consisting of C₅₋₃₀ alkyl, C₅₋₂₀ alkenyl, —R*YR″, —YR″, and —R″M′R′;

R₂ and R₃ are independently selected from the group consisting of H, C₁₋₁₄ alkyl, C₂₋₁₄ alkenyl, —R*YR″, —YR″, and —R*OR″, or R₂ and R₃, together with the atom to which they are attached, form a heterocycle or carbocycle;

R₄ is selected from the group consisting of a C₃₋₆ carbocycle, —(CH₂)_(n)Q, —(CH₂)_(n)CHQR, —CHQR, —CQ(R)₂, and unsubstituted C₁₋₆ alkyl, where Q is selected from a C₃₋₆ carbocycle, a 5- to 14-membered heteroaryl having one or more heteroatoms selected from N, O, and S, —OR,

—O(CH₂)_(n)N(R)₂, —C(O)OR, —OC(O)R, —CX₃, —CX₂H, —CXH₂, —CN, —C(O)N(R)₂, —N(R)C(O)R, —N(R)S(O)₂R, —N(R)C(O)N(R)₂, —N(R)C(S)N(R)₂, —CRN(R)₂C(O)OR, —N(R)R₈, —O(CH₂)_(n)OR, —N(R)C(═NR₉)N(R)₂, —N(R)C(═CHR₉)N(R)₂, —OC(O)N(R)₂, —N(R)C(O)OR, —N(OR)C(O)R, —N(OR)S(O)₂R, —N(OR)C(O)OR, —N(OR)C(O)N(R)₂, —N(OR)C(S)N(R)₂, —N(OR)C(═NR₉)N(R)₂, —N(OR)C(═CHR₉)N(R)₂, —C(═NR₉)N(R)₂, —C(═NR₉)R, —C(O)N(R)OR, and a 5- to 14-membered heterocycloalkyl having one or more heteroatoms selected from N, O, and S which is substituted with one or more substituents selected from oxo (═O), OH, amino, mono- or di-alkylamino, and C₁₋₃ alkyl, and each n is independently selected from 1, 2, 3, 4, and 5;

each R₅ is independently selected from the group consisting of C₁₋₃ alkyl, C₂₋₃ alkenyl, and H;

each R₆ is independently selected from the group consisting of C₁₋₃ alkyl, C₂₋₃ alkenyl, and H;

M and M′ are independently selected from —C(O)O—, —OC(O)—, —C(O)N(R′)—, —N(R′)C(O)—, —C(O)—, —C(S)—, —C(S)S—, —SC(S)—, —CH(OH)—, —P(O)(OR′)O—, —S(O)₂—, —S—S—, an aryl group, and a heteroaryl group;

R₇ is selected from the group consisting of C₁₋₃ alkyl, C₂₋₃ alkenyl, and H;

R₈ is selected from the group consisting of C₃₋₆ carbocycle and heterocycle;

R₉ is selected from the group consisting of H, CN, NO₂, C₁₋₆ alkyl, —OR, —S(O)₂R, —S(O)₂N(R)₂, C₂₋₆ alkenyl, C₃₋₆ carbocycle and heterocycle;

each R is independently selected from the group consisting of C₁₋₃ alkyl, C₂₋₃ alkenyl, and H;

each R′ is independently selected from the group consisting of C₁₋₁₈ alkyl, C₂₋₁₈ alkenyl, —R*YR″, —YR″, and H;

each R″ is independently selected from the group consisting of C₃₋₁₄ alkyl and C₃₋₁₄ alkenyl;

each R* is independently selected from the group consisting of C₁₋₁₂ alkyl and C₂₋₁₂ alkenyl;

each Y is independently a C₃₋₆ carbocycle;

each X is independently selected from the group consisting of F, Cl, Br, and I; and

m is selected from 5, 6, 7, 8, 9, 10, 11, 12, and 13,

or salts or isomers thereof.

In some embodiments, another subset of compounds of Formula (I) includes those in which

R₁ is selected from the group consisting of C₅₋₃₀ alkyl, C₅₋₂₀ alkenyl, —R*YR″, —YR″, and —R″M′R′;

R₂ and R₃ are independently selected from the group consisting of H, C₁₋₁₄ alkyl, C₂₋₁₄ alkenyl, —R*YR″, —YR″, and —R*OR″, or R₂ and R₃, together with the atom to which they are attached, form a heterocycle or carbocycle;

R₄ is selected from the group consisting of a C₃₋₆ carbocycle, —(CH₂)_(n)Q, —(CH₂)_(n)CHQR, —CHQR, —CQ(R)₂, and unsubstituted C₁₋₆ alkyl, where Q is selected from a C₃₋₆ carbocycle, a 5- to 14-membered heterocycle having one or more heteroatoms selected from N, O, and S, —OR,

—O(CH₂)_(n)N(R)₂, —C(O)OR, —OC(O)R, —CX₃, —CX₂H, —CXH₂, —CN, —C(O)N(R)₂, —N(R)C(O)R, —N(R)S(O)₂R, —N(R)C(O)N(R)₂, —N(R)C(S)N(R)₂, —CRN(R)₂C(O)OR, —N(R)R₈, —O(CH₂)_(n)OR, —N(R)C(═NR₉)N(R)₂, —N(R)C(═CHR₉)N(R)₂, —OC(O)N(R)₂, —N(R)C(O)OR, —N(OR)C(O)R, —N(OR)S(O)₂R, —N(OR)C(O)OR, —N(OR)C(O)N(R)₂, —N(OR)C(S)N(R)₂, —N(OR)C(═NR₉)N(R)₂, —N(OR)C(═CHR₉)N(R)₂, —C(═NR₉)R, —C(O)N(R)OR, and —C(═NR₉)N(R)₂, and each n is independently selected from 1, 2, 3, 4, and 5; and when Q is a 5- to 14-membered heterocycle and (i) R₄ is —(CH₂)_(n)Q in which n is 1 or 2, or (ii) R₄ is —(CH₂)_(n)CHQR in which n is 1, or (iii) R₄ is —CHQR, and —CQ(R)₂, then Q is either a 5- to 14-membered heteroaryl or 8- to 14-membered heterocycloalkyl;

each R₅ is independently selected from the group consisting of C₁₋₃ alkyl, C₂₋₃ alkenyl, and H;

each R₆ is independently selected from the group consisting of C₁₋₃ alkyl, C₂₋₃ alkenyl, and H;

M and M′ are independently selected from —C(O)O—, —OC(O)—, —C(O)N(R′)—, —N(R′)C(O)—, —C(O)—, —C(S)—, —C(S)S—, —SC(S)—, —CH(OH)—, —P(O)(OR′)O—, —S(O)₂—, —S—S—, an aryl group, and a heteroaryl group;

R₇ is selected from the group consisting of C₁₋₃ alkyl, C₂₋₃ alkenyl, and H;

R₈ is selected from the group consisting of C₃₋₆ carbocycle and heterocycle;

R₉ is selected from the group consisting of H, CN, NO₂, C₁₋₆ alkyl, —OR, —S(O)₂R, —S(O)₂N(R)₂, C₂₋₆ alkenyl, C₃₋₆ carbocycle and heterocycle;

each R is independently selected from the group consisting of C₁₋₃ alkyl, C₂₋₃ alkenyl, and H;

each R′ is independently selected from the group consisting of C₁₋₁₈ alkyl, C₂₋₁₈ alkenyl, —R*YR″, —YR″, and H;

each R″ is independently selected from the group consisting of C₃₋₁₄ alkyl and C₃₋₁₄ alkenyl;

each R* is independently selected from the group consisting of C₁₋₁₂ alkyl and C₂₋₁₂ alkenyl;

each Y is independently a C₃₋₆ carbocycle;

each X is independently selected from the group consisting of F, Cl, Br, and I; and

m is selected from 5, 6, 7, 8, 9, 10, 11, 12, and 13, or salts or isomers thereof.

In some embodiments, another subset of compounds of Formula (I) includes those in which

R₁ is selected from the group consisting of C₅₋₃₀ alkyl, C₅₋₂₀ alkenyl, —R*YR″, —YR″, and —R″M′R′;

R₂ and R₃ are independently selected from the group consisting of H, C₁₋₁₄ alkyl, C₂₋₁₄ alkenyl, —R*YR″, —YR″, and —R*OR″, or R₂ and R₃, together with the atom to which they are attached, form a heterocycle or carbocycle;

R₄ is selected from the group consisting of a C₃₋₆ carbocycle, —(CH₂)_(n)Q, —(CH₂)_(n)CHQR, —CHQR, —CQ(R)₂, and unsubstituted C₁₋₆ alkyl, where Q is selected from a C₃₋₆ carbocycle, a 5- to 14-membered heteroaryl having one or more heteroatoms selected from N, O, and S, —OR,

—O(CH₂)_(n)N(R)₂, —C(O)OR, —OC(O)R, —CX₃, —CX₂H, —CXH₂, —CN, —C(O)N(R)₂, —N(R)C(O)R, —N(R)S(O)₂R, —N(R)C(O)N(R)₂, —N(R)C(S)N(R)₂, —CRN(R)₂C(O)OR, —N(R)R₈, —O(CH₂)_(n)OR, —N(R)C(═NR₉)N(R)₂, —N(R)C(═CHR₉)N(R)₂, —OC(O)N(R)₂, —N(R)C(O)OR, —N(OR)C(O)R, —N(OR)S(O)₂R, —N(OR)C(O)OR, —N(OR)C(O)N(R)₂, —N(OR)C(S)N(R)₂, —N(OR)C(═NR₉)N(R)₂, —N(OR)C(═CHR₉)N(R)₂, —C(═NR₉)R, —C(O)N(R)OR, and —C(═NR₉)N(R)₂, and each n is independently selected from 1, 2, 3, 4, and 5;

each R₅ is independently selected from the group consisting of C₁₋₃ alkyl, C₂₋₃ alkenyl, and H;

each R₆ is independently selected from the group consisting of C₁₋₃ alkyl, C₂₋₃ alkenyl, and H;

M and M′ are independently selected from —C(O)O—, —OC(O)—, —C(O)N(R′)—, —N(R′)C(O)—, —C(O)—, —C(S)—, —C(S)S—, —SC(S)—, —CH(OH)—, —P(O)(OR′)O—, —S(O)₂—, —S—S—, an aryl group, and a heteroaryl group;

R₇ is selected from the group consisting of C₁₋₃ alkyl, C₂₋₃ alkenyl, and H;

R₈ is selected from the group consisting of C₃₋₆ carbocycle and heterocycle;

R₉ is selected from the group consisting of H, CN, NO₂, C₁₋₆ alkyl, —OR, —S(O)₂R, —S(O)₂N(R)₂, C₂₋₆ alkenyl, C₃₋₆ carbocycle and heterocycle;

each R is independently selected from the group consisting of C₁₋₃ alkyl, C₂₋₃ alkenyl, and H;

each R′ is independently selected from the group consisting of C₁₋₁₈ alkyl, C₂₋₁₈ alkenyl, —R*YR″, —YR″, and H;

each R″ is independently selected from the group consisting of C₃₋₁₄ alkyl and C₃₋₁₄ alkenyl;

each R* is independently selected from the group consisting of C₁₋₁₂ alkyl and C₂₋₁₂ alkenyl;

each Y is independently a C₃₋₆ carbocycle;

each X is independently selected from the group consisting of F, Cl, Br, and I; and

m is selected from 5, 6, 7, 8, 9, 10, 11, 12, and 13, or salts or isomers thereof.

In some embodiments, another subset of compounds of Formula (I) includes those in which

R₁ is selected from the group consisting of C₅₋₃₀ alkyl, C₅₋₂₀ alkenyl, —R*YR″, —YR″, and —R″M′R′;

R₂ and R₃ are independently selected from the group consisting of H, C₂₋₁₄ alkyl, C₂₋₁₄ alkenyl, —R*YR″, —YR″, and —R*OR″, or R₂ and R₃, together with the atom to which they are attached, form a heterocycle or carbocycle;

R₄ is —(CH₂)_(n)Q or —(CH₂)_(n)CHQR, where Q is —N(R)₂, and n is selected from 3, 4, and 5;

each R₅ is independently selected from the group consisting of C₁₋₃ alkyl, C₂₋₃ alkenyl, and H;

each R₆ is independently selected from the group consisting of C₁₋₃ alkyl, C₂₋₃ alkenyl, and H;

M and M′ are independently selected from —C(O)O—, —OC(O)—, —C(O)N(R′)—, —N(R′)C(O)—, —C(O)—, —C(S)—, —C(S)S—, —SC(S)—, —CH(OH)—, —P(O)(OR′)O—, —S(O)₂—, —S—S—, an aryl group, and a heteroaryl group;

R₇ is selected from the group consisting of C₁₋₃ alkyl, C₂₋₃ alkenyl, and H;

each R is independently selected from the group consisting of C₁₋₃ alkyl, C₂₋₃ alkenyl, and H;

each R′ is independently selected from the group consisting of C₁₋₁₈ alkyl, C₂₋₁₈ alkenyl, —R*YR″, —YR″, and H;

each R″ is independently selected from the group consisting of C₃₋₁₄ alkyl and C₃₋₁₄ alkenyl;

each R* is independently selected from the group consisting of C₁₋₁₂ alkyl and C₁₋₁₂ alkenyl;

each Y is independently a C₃₋₆ carbocycle;

each X is independently selected from the group consisting of F, Cl, Br, and I; and

m is selected from 5, 6, 7, 8, 9, 10, 11, 12, and 13,

or salts or isomers thereof.

In some embodiments, another subset of compounds of Formula (I) includes those in which

R₁ is selected from the group consisting of C₅₋₃₀ alkyl, C₅₋₂₀ alkenyl, —R*YR″, —YR″, and —R″M′R′;

R₂ and R₃ are independently selected from the group consisting of C₁₋₁₄ alkyl, C₂₋₁₄ alkenyl, —R*YR″, —YR″, and —R*OR″, or R₂ and R₃, together with the atom to which they are attached, form a heterocycle or carbocycle;

R₄ is selected from the group consisting of —(CH₂)_(n)Q, —(CH₂)_(n)CHQR, —CHQR, and —CQ(R)₂, where Q is —N(R)₂, and n is selected from 1, 2, 3, 4, and 5;

each R₅ is independently selected from the group consisting of C₁₋₃ alkyl, C₂₋₃ alkenyl, and H;

each R₆ is independently selected from the group consisting of C₁₋₃ alkyl, C₂₋₃ alkenyl, and H;

M and M′ are independently selected from —C(O)O—, —OC(O)—, —C(O)N(R′)—, —N(R′)C(O)—, —C(O)—, —C(S)—, —C(S)S—, —SC(S)—, —CH(OH)—, —P(O)(OR′)O—, —S(O)₂—, —S—S—, an aryl group, and a heteroaryl group;

R₇ is selected from the group consisting of C₁₋₃ alkyl, C₂₋₃ alkenyl, and H;

each R is independently selected from the group consisting of C₁₋₃ alkyl, C₂₋₃ alkenyl, and H;

each R′ is independently selected from the group consisting of C₁₋₁₈ alkyl, C₂₋₁₈ alkenyl, —R*YR″, —YR″, and H;

each R″ is independently selected from the group consisting of C₃₋₁₄ alkyl and C₃₋₁₄ alkenyl;

each R* is independently selected from the group consisting of C₁₋₁₂ alkyl and C₁₋₁₂ alkenyl;

each Y is independently a C₃₋₆ carbocycle;

each X is independently selected from the group consisting of F, Cl, Br, and I; and

m is selected from 5, 6, 7, 8, 9, 10, 11, 12, and 13,

or salts or isomers thereof.

In some embodiments, a subset of compounds of Formula (I) includes those of Formula (IA):

or a salt or isomer thereof, wherein 1 is selected from 1, 2, 3, 4, and 5; m is selected from 5, 6, 7, 8, and 9; M₁ is a bond or M′; R₄ is unsubstituted C₁₋₃ alkyl, or —(CH₂)_(n)Q, in which Q is OH, —NHC(S)N(R)₂, —NHC(O)N(R)₂, —N(R)C(O)R, —N(R)S(O)₂R, —N(R)R₈, —NHC(═NR₉)N(R)₂, —NHC(═CHR₉)N(R)₂, —OC(O)N(R)₂, —N(R)C(O)OR, heteroaryl or heterocycloalkyl; M and M′ are independently selected

from —C(O)O—, —OC(O)—, —C(O)N(R′)—, —P(O)(OR′)O—, —S—S—, an aryl group, and a heteroaryl group; and R₂ and R₃ are independently selected from the group consisting of H, C₁₋₁₄ alkyl, and C₂₋₁₄ alkenyl.

In some embodiments, a subset of compounds of Formula (I) includes those of Formula (II):

or a salt or isomer thereof, wherein 1 is selected from 1, 2, 3, 4, and 5; M₁ is a bond or M′; R₄ is unsubstituted C₁₋₃ alkyl, or —(CH₂)_(n)Q, in which n is 2, 3, or 4, and Q is OH, —NHC(S)N(R)₂, —NHC(O)N(R)₂, —N(R)C(O)R, —N(R)S(O)₂R, —N(R)R₈, —NHC(═NR₉)N(R)₂, —NHC(═CHR₉)N(R)₂, —OC(O)N(R)₂, —N(R)C(O)OR, heteroaryl or heterocycloalkyl; M and M′ are independently selected from —C(O)O—, —OC(O)—, —C(O)N(R′)—, —P(O)(OR′)O—, —S—S—, an aryl group, and a heteroaryl group; and R₂ and R₃ are independently selected from the group consisting of H, C₁₋₁₄ alkyl, and C₂₋₁₄ alkenyl.

In some embodiments, a subset of compounds of Formula (I) includes those of Formula (IIa), (IIb), (IIc), or (IIe):

or a salt or isomer thereof, wherein R₄ is as described herein.

In some embodiments, a subset of compounds of Formula (I) includes those of Formula (IId):

or a salt or isomer thereof, wherein n is 2, 3, or 4; and m, R′, R″, and R₂ through R₆ are as described herein. For example, each of R₂ and R₃ may be independently selected from the group consisting of C₅₋₁₄ alkyl and C₅₋₁₄ alkenyl.

In some embodiments, the compound of Formula (I) is selected from the group consisting of:

In further embodiments, the compound of Formula (I) is selected from the group consisting of:

In some embodiments, the compound of Formula (I) is selected from the group consisting of:

and salts and isomers thereof.

In some embodiments, a nanoparticle comprises the following compound:

or salts and isomers thereof.

In some embodiments, the disclosure features a nanoparticle composition including a lipid component comprising a compound as described herein (e.g., a compound according to Formula (I), (IA), (II), (IIa), (IIb), (IIc), (IId) or (IIe)).

In some embodiments, the disclosure features a pharmaceutical composition comprising a nanoparticle composition according to the preceding embodiments and a pharmaceutically acceptable carrier. For example, the pharmaceutical composition is refrigerated or frozen for storage and/or shipment (e.g., being stored at a temperature of 4° C. or lower, such as a temperature between about −150° C. and about 0° C. or between about −80° C. and about −20° C. (e.g., about −5° C., −10° C., −15° C., −20° C., −25° C., −30° C., −40° C., −50° C., −60° C., −70° C., −80° C., −90° C., −130° C. or −150° C.). For example, the pharmaceutical composition is a solution that is refrigerated for storage and/or shipment at, for example, about −20° C., −30° C., −40° C., −50° C., −60° C., −70° C., or −80° C.

In some embodiments, the disclosure provides a method of delivering a therapeutic and/or prophylactic (e.g., RNA, such as mRNA) to a cell (e.g., a mammalian cell). This method includes the step of administering to a subject (e.g., a mammal, such as a human) a nanoparticle composition including (i) a lipid component including a phospholipid (such as a polyunsaturated lipid), a PEG lipid, a structural lipid, and a compound of Formula (I), (IA), (II), (IIa), (IIb), (IIc), (IId) or (IIe) and (ii) a therapeutic and/or prophylactic, in which administering involves contacting the cell with the nanoparticle composition, whereby the therapeutic and/or prophylactic is delivered to the cell.

In some embodiments, the disclosure provides a method of producing a polypeptide of interest in a cell (e.g., a mammalian cell). The method includes the step of contacting the cell with a nanoparticle composition including (i) a lipid component including a phospholipid (such as a polyunsaturated lipid), a PEG lipid, a structural lipid, and a compound of Formula (I), (IA), (II), (IIa), (IIb), (IIc), (IId) or (IIe) and (ii) an mRNA encoding the polypeptide of interest, whereby the mRNA is capable of being translated in the cell to produce the polypeptide.

In some embodiments, the disclosure provides a method of treating a disease or disorder in a mammal (e.g., a human) in need thereof. The method includes the step of administering to the mammal a therapeutically effective amount of a nanoparticle composition including (i) a lipid component including a phospholipid (such as a polyunsaturated lipid), a PEG lipid, a structural lipid, and a compound of Formula (I), (IA), (II), (IIa), (IIb), (IIc), (IId) or (IIe) and (ii) a therapeutic and/or prophylactic (e.g., an mRNA). In some embodiments, the disease or disorder is characterized by dysfunctional or aberrant protein or polypeptide activity. For example, the disease or disorder is selected from the group consisting of rare diseases, infectious diseases, cancer and proliferative diseases, genetic diseases (e.g., cystic fibrosis), autoimmune diseases, diabetes, neurodegenerative diseases, cardio- and reno-vascular diseases, and metabolic diseases.

In some embodiments, the disclosure provides a method of delivering (e.g., specifically delivering) a therapeutic and/or prophylactic to a mammalian organ (e.g., a liver, spleen, lung, or femur). This method includes the step of administering to a subject (e.g., a mammal) a nanoparticle composition including (i) a lipid component including a phospholipid, a PEG lipid, a structural lipid, and a compound of Formula (I), (IA), (II), (IIa), (IIb), (IIc), (IId) or (IIe) and (ii) a therapeutic and/or prophylactic (e.g., an mRNA), in which administering involves contacting the cell with the nanoparticle composition, whereby the therapeutic and/or prophylactic is delivered to the target organ (e.g., a liver, spleen, lung, or femur).

In some embodiments, the disclosure features a method for the enhanced delivery of a therapeutic and/or prophylactic (e.g., an mRNA) to a target tissue (e.g., a liver, spleen, lung, or femur). This method includes administering to a subject (e.g., a mammal) a nanoparticle composition, the composition including (i) a lipid component including a compound of Formula (I), (IA), (II), (IIa), (IIb), (IIc), (IId) or (IIe), a phospholipid, a structural lipid, and a PEG lipid; and (ii) a therapeutic and/or prophylactic, the administering including contacting the target tissue with the nanoparticle composition, whereby the therapeutic and/or prophylactic is delivered to the target tissue.

In some embodiments, the disclosure features a method of lowering immunogenicity comprising introducing the nanoparticle composition of the disclosure into cells, wherein the nanoparticle composition reduces the induction of the cellular immune response of the cells to the nanoparticle composition, as compared to the induction of the cellular immune response in cells induced by a reference composition which comprises a reference lipid instead of a compound of Formula (I), (IA), (II), (IIa), (IIb), (IIc), (IId) or (IIe). For example, the cellular immune response is an innate immune response, an adaptive immune response, or both.

The disclosure also includes methods of synthesizing a compound of Formula (I), (IA), (II), (IIa), (IIb), (IIc), (IId) or (IIe) and methods of making a nanoparticle composition including a lipid component comprising the compound of Formula (I), (IA), (II), (IIa), (IIb), (IIc), (IId) or (IIe).

Modes of Vaccine Administration

Tropical disease RNA (e.g. mRNA) vaccines may be administered by any route which results in a therapeutically effective outcome. These include, but are not limited, to intradermal, intramuscular, intranasal and/or subcutaneous administration. The present disclosure provides methods comprising administering RNA (e.g., mRNA) vaccines to a subject in need thereof. The exact amount required will vary from subject to subject, depending on the species, age, and general condition of the subject, the severity of the disease, the particular composition, its mode of administration, its mode of activity, and the like. Tropical disease RNA (e.g., mRNA) vaccines compositions are typically formulated in dosage unit form for ease of administration and uniformity of dosage. It will be understood, however, that the total daily usage of RNA (e.g., mRNA) vaccine compositions may be decided by the attending physician within the scope of sound medical judgment. The specific therapeutically effective, prophylactically effective, or appropriate imaging dose level for any particular patient will depend upon a variety of factors including the disorder being treated and the severity of the disorder; the activity of the specific compound employed; the specific composition employed; the age, body weight, general health, sex and diet of the patient; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific compound employed; and like factors well known in the medical arts.

In some embodiments, tropical disease RNA (e.g. mRNA) vaccines compositions may be administered at dosage levels sufficient to deliver 0.0001 mg/kg to 100 mg/kg, 0.001 mg/kg to 0.05 mg/kg, 0.005 mg/kg to 0.05 mg/kg, 0.001 mg/kg to 0.005 mg/kg, 0.05 mg/kg to 0.5 mg/kg, 0.01 mg/kg to 50 mg/kg, 0.1 mg/kg to 40 mg/kg, 0.5 mg/kg to 30 mg/kg, 0.01 mg/kg to 10 mg/kg, 0.1 mg/kg to 10 mg/kg, or 1 mg/kg to 25 mg/kg, of subject body weight per day, one or more times a day, per week, per month, etc. to obtain the desired therapeutic, diagnostic, prophylactic, or imaging effect (see, e.g., the range of unit doses described in International Publication No WO2013078199, the contents of which are herein incorporated by reference in their entirety). The desired dosage may be delivered three times a day, two times a day, once a day, every other day, every third day, every week, every two weeks, every three weeks, every four weeks, every 2 months, every three months, every 6 months, etc. In some embodiments, the desired dosage may be delivered using multiple administrations (e.g., two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, or more administrations). When multiple administrations are employed, split dosing regimens such as those described herein may be used. In exemplary embodiments, tropical disease RNA (e.g., mRNA) vaccines compositions may be administered at dosage levels sufficient to deliver 0.0005 mg/kg to 0.01 mg/kg, e.g., about 0.0005 mg/kg to about 0.0075 mg/kg, e.g., about 0.0005 mg/kg, about 0.001 mg/kg, about 0.002 mg/kg, about 0.003 mg/kg, about 0.004 mg/kg or about 0.005 mg/kg.

In some embodiments, tropical disease RNA (e.g., mRNA) vaccine compositions may be administered once or twice (or more) at dosage levels sufficient to deliver 0.025 mg/kg to 0.250 mg/kg, 0.025 mg/kg to 0.500 mg/kg, 0.025 mg/kg to 0.750 mg/kg, or 0.025 mg/kg to 1.0 mg/kg.

In some embodiments, tropical disease RNA (e.g., mRNA) vaccine compositions may be administered twice (e.g., Day 0 and Day 7, Day 0 and Day 14, Day 0 and Day 21, Day 0 and Day 28, Day 0 and Day 60, Day 0 and Day 90, Day 0 and Day 120, Day 0 and Day 150, Day 0 and Day 180, Day 0 and 3 months later, Day 0 and 6 months later, Day 0 and 9 months later, Day 0 and 12 months later, Day 0 and 18 months later, Day 0 and 2 years later, Day 0 and 5 years later, or Day 0 and 10 years later) at a total dose of or at dosage levels sufficient to deliver a total dose of 0.0100 mg, 0.025 mg, 0.050 mg, 0.075 mg, 0.100 mg, 0.125 mg, 0.150 mg, 0.175 mg, 0.200 mg, 0.225 mg, 0.250 mg, 0.275 mg, 0.300 mg, 0.325 mg, 0.350 mg, 0.375 mg, 0.400 mg, 0.425 mg, 0.450 mg, 0.475 mg, 0.500 mg, 0.525 mg, 0.550 mg, 0.575 mg, 0.600 mg, 0.625 mg, 0.650 mg, 0.675 mg, 0.700 mg, 0.725 mg, 0.750 mg, 0.775 mg, 0.800 mg, 0.825 mg, 0.850 mg, 0.875 mg, 0.900 mg, 0.925 mg, 0.950 mg, 0.975 mg, or 1.0 mg. Higher and lower dosages and frequency of administration are encompassed by the present disclosure. For example, a tropical disease RNA (e.g., mRNA) vaccine composition may be administered three or four times.

In some embodiments, tropical disease RNA (e.g., mRNA) vaccine compositions may be administered twice (e.g., Day 0 and Day 7, Day 0 and Day 14, Day 0 and Day 21, Day 0 and Day 28, Day 0 and Day 60, Day 0 and Day 90, Day 0 and Day 120, Day 0 and Day 150, Day 0 and Day 180, Day 0 and 3 months later, Day 0 and 6 months later, Day 0 and 9 months later, Day 0 and 12 months later, Day 0 and 18 months later, Day 0 and 2 years later, Day 0 and 5 years later, or Day 0 and 10 years later) at a total dose of or at dosage levels sufficient to deliver a total dose of 0.010 mg, 0.025 mg, 0.100 mg or 0.400 mg.

In some embodiments, the tropical disease RNA (e.g., mRNA) vaccine for use in a method of vaccinating a subject is administered to the subject as a single dosage of between 10 μg/kg and 400 μg/kg of the nucleic acid vaccine (in an effective amount to vaccinate the subject). In some embodiments the RNA (e.g., mRNA) vaccine for use in a method of vaccinating a subject is administered to the subject as a single dosage of between 10 μg and 400 μg of the nucleic acid vaccine (in an effective amount to vaccinate the subject). In some embodiments, a tropical disease RNA (e.g., mRNA) vaccine for use in a method of vaccinating a subject is administered to the subject as a single dosage of 25-1000 μg (e.g., a single dosage of mRNA encoding Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigen). In some embodiments, a tropical disease RNA (e.g., mRNA) vaccine is administered to the subject as a single dosage of 25, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950 or 1000 μg. For example, a tropical disease RNA (e.g., mRNA) vaccine may be administered to a subject as a single dose of 25-100, 25-500, 50-100, 50-500, 50-1000, 100-500, 100-1000, 250-500, 250-1000, or 500-1000 μg. In some embodiments, a tropical disease RNA (e.g., mRNA) vaccine for use in a method of vaccinating a subject is administered to the subject as two dosages, the combination of which equals 25-1000 μg of the tropical disease RNA (e.g., mRNA) vaccine.

A tropical disease RNA (e.g. mRNA) vaccine pharmaceutical composition described herein can be formulated into a dosage form described herein, such as an intranasal, intratracheal, or injectable (e.g., intravenous, intraocular, intravitreal, intramuscular, intradermal, intracardiac, intraperitoneal, intranasal and subcutaneous).

Tropical Disease RNA (e.g., mRNA) Vaccine Formulations and Methods of Use

Some aspects of the present disclosure provide formulations of the tropical disease RNA (e.g., mRNA) vaccine, wherein the RNA (e.g., mRNA) vaccine is formulated in an effective amount to produce an antigen specific immune response in a subject (e.g., production of antibodies specific to an Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide). An “effective amount” is a dose of an RNA (e.g., mRNA) vaccine effective to produce an antigen-specific immune response. Also provided herein are methods of inducing an antigen-specific immune response in a subject.

In some embodiments, the antigen-specific immune response is characterized by measuring an anti-Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide antibody titer produced in a subject administered a tropical disease RNA (e.g., mRNA) vaccine as provided herein. An antibody titer is a measurement of the amount of antibodies within a subject, for example, antibodies that are specific to a particular antigen (e.g., an Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide) or epitope of an antigen. Antibody titer is typically expressed as the inverse of the greatest dilution that provides a positive result. Enzyme-linked immunosorbent assay (ELISA) is a common assay for determining antibody titers, for example.

In some embodiments, an antibody titer is used to assess whether a subject has had an infection or to determine whether immunizations are required. In some embodiments, an antibody titer is used to determine the strength of an autoimmune response, to determine whether a booster immunization is needed, to determine whether a previous vaccine was effective, and to identify any recent or prior infections. In accordance with the present disclosure, an antibody titer may be used to determine the strength of an immune response induced in a subject by the tropical disease RNA (e.g., mRNA) vaccine.

In some embodiments, an anti-antigenic polypeptide (e.g., an anti-Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide) antibody titer produced in a subject is increased by at least 1 log relative to a control. For example, anti-antigenic polypeptide antibody titer produced in a subject may be increased by at least 1.5, at least 2, at least 2.5, or at least 3 log relative to a control. In some embodiments, the anti-antigenic polypeptide antibody titer produced in the subject is increased by 1, 1.5, 2, 2.5 or 3 log relative to a control. In some embodiments, the anti-antigenic polypeptide antibody titer produced in the subject is increased by 1-3 log relative to a control. For example, the anti-antigenic polypeptide antibody titer produced in a subject may be increased by 1-1.5, 1-2, 1-2.5, 1-3, 1.5-2, 1.5-2.5, 1.5-3, 2-2.5, 2-3, or 2.5-3 log relative to a control.

In some embodiments, the anti-antigenic polypeptide (e.g., an anti-Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide) antibody titer produced in a subject is increased at least 2 times relative to a control. For example, the anti-antigenic polypeptide antibody titer produced in a subject may be increased at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, or at least 10 times relative to a control. In some embodiments, the anti-antigenic polypeptide antibody titer produced in the subject is increased 2, 3, 4, 5, 6, 7, 8, 9, or 10 times relative to a control. In some embodiments, the anti-antigenic polypeptide antibody titer produced in a subject is increased 2-10 times relative to a control. For example, the anti-antigenic polypeptide antibody titer produced in a subject may be increased 2-10, 2-9, 2-8, 2-7, 2-6, 2-5, 2-4, 2-3, 3-10, 3-9, 3-8, 3-7, 3-6, 3-5, 3-4, 4-10, 4-9, 4-8, 4-7, 4-6, 4-5, 5-10, 5-9, 5-8, 5-7, 5-6, 6-10, 6-9, 6-8, 6-7, 7-10, 7-9, 7-8, 8-10, 8-9, or 9-10 times relative to a control.

A control, in some embodiments, is the anti-antigenic polypeptide (e.g., an anti-Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide) antibody titer produced in a subject who has not been administered a tropical disease RNA (e.g., mRNA) vaccine of the present disclosure. In some embodiments, a control is an anti-antigenic polypeptide (e.g., an anti-Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide) antibody titer produced in a subject who has been administered a live attenuated Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV vaccine. An attenuated vaccine is a vaccine produced by reducing the virulence of a viable (live) virus. An attenuated virus is altered in a manner that renders it harmless or less virulent relative to a live, unmodified virus. In some embodiments, a control is an anti-antigenic polypeptide (e.g., an anti-Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide) antibody titer produced in a subject administered inactivated Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV vaccine. In some embodiments, a control is an anti-antigenic polypeptide (e.g., an anti-Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide) antibody titer produced in a subject administered a recombinant or purified Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV protein vaccine. Recombinant protein vaccines typically include protein antigens that either have been produced in a heterologous expression system (e.g., bacteria or yeast) or purified from large amounts of the pathogenic organism. In some embodiments, a control is an anti-antigenic polypeptide (e.g., an anti-Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide) antibody titer produced in a subject who has been administered an Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV virus-like particle (VLP) vaccine.

In some embodiments, an effective amount of a tropical disease RNA (e.g., mRNA) vaccine is a dose that is reduced compared to the standard of care dose of a recombinant Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV protein vaccine. A “standard of care,” as provided herein, refers to a medical or psychological treatment guideline and can be general or specific. “Standard of care” specifies appropriate treatment based on scientific evidence and collaboration between medical professionals involved in the treatment of a given condition. It is the diagnostic and treatment process that a physician/clinician should follow for a certain type of patient, illness or clinical circumstance. A “standard of care dose,” as provided herein, refers to the dose of a recombinant or purified Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV protein vaccine, or a live attenuated or inactivated Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV vaccine, that a physician/clinician or other medical professional would administer to a subject to treat or prevent Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV, or a related condition, while following the standard of care guideline for treating or preventing Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV, or a related condition.

In some embodiments, the anti-antigenic polypeptide (e.g., an anti-Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide) antibody titer produced in a subject administered an effective amount of a tropical disease RNA (e.g., mRNA) vaccine is equivalent to an anti-antigenic polypeptide (e.g., an anti-Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptide) antibody titer produced in a control subject administered a standard of care dose of a recombinant or purified Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV protein vaccine or a live attenuated or inactivated Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV vaccine.

In some embodiments, an effective amount of a tropical disease RNA (e.g., mRNA) vaccine is a dose equivalent to an at least 2-fold reduction in a standard of care dose of a recombinant or purified Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV protein vaccine. For example, an effective amount of a tropical disease RNA (e.g., mRNA) vaccine may be a dose equivalent to an at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, or at least 10-fold reduction in a standard of care dose of a recombinant or purified Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV protein vaccine. In some embodiments, an effective amount of a tropical disease RNA (e.g., mRNA) vaccine is a dose equivalent to an at least at least 100-fold, at least 500-fold, or at least 1000-fold reduction in a standard of care dose of a recombinant or purified Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV protein vaccine. In some embodiments, an effective amount of a tropical disease RNA (e.g., mRNA) vaccine is a dose equivalent to a 2-, 3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, 20-, 50-, 100-, 250-, 500-, or 1000-fold reduction in a standard of care dose of a recombinant or purified Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV protein vaccine. In some embodiments, the anti-antigenic polypeptide antibody titer produced in a subject administered an effective amount of a tropical disease RNA (e.g., mRNA) vaccine is equivalent to an anti-antigenic polypeptide antibody titer produced in a control subject administered the standard of care dose of a recombinant or protein Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV protein vaccine or a live attenuated or inactivated Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV vaccine. In some embodiments, an effective amount of a tropical disease RNA (e.g., mRNA) vaccine is a dose equivalent to a 2-fold to 1000-fold (e.g., 2-fold to 100-fold, 10-fold to 1000-fold) reduction in the standard of care dose of a recombinant or purified Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV protein vaccine, wherein the anti-antigenic polypeptide antibody titer produced in the subject is equivalent to an anti-antigenic polypeptide antibody titer produced in a control subject administered the standard of care dose of a recombinant or purified Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV protein vaccine or a live attenuated or inactivated Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV vaccine.

In some embodiments, the effective amount of a tropical disease RNA (e.g., mRNA) vaccine is a dose equivalent to a 2 to 1000-, 2 to 900-, 2 to 800-, 2 to 700-, 2 to 600-, 2 to 500-, 2 to 400-, 2 to 300-, 2 to 200-, 2 to 100-, 2 to 90-, 2 to 80-, 2 to 70-, 2 to 60-, 2 to 50-, 2 to 40-, 2 to 30-, 2 to 20-, 2 to 10-, 2 to 9-, 2 to 8-, 2 to 7-, 2 to 6-, 2 to 5-, 2 to 4-, 2 to 3-, 3 to 1000-, 3 to 900-, 3 to 800-, 3 to 700-, 3 to 600-, 3 to 500-, 3 to 400-, 3 to 3 to 00-, 3 to 200-, 3 to 100-, 3 to 90-, 3 to 80-, 3 to 70-, 3 to 60-, 3 to 50-, 3 to 40-, 3 to 30-, 3 to 20-, 3 to 10-, 3 to 9-, 3 to 8-, 3 to 7-, 3 to 6-, 3 to 5-, 3 to 4-, 4 to 1000-, 4 to 900-, 4 to 800-, 4 to 700-, 4 to 600-, 4 to 500-, 4 to 400-, 4 to 4 to 00-, 4 to 200-, 4 to 100-, 4 to 90-, 4 to 80-, 4 to 70-, 4 to 60-, 4 to 50-, 4 to 40-, 4 to 30-, 4 to 20-, 4 to 10-, 4 to 9-, 4 to 8-, 4 to 7-, 4 to 6-, 4 to 5-, 4 to 4-, 5 to 1000-, 5 to 900-, 5 to 800-, 5 to 700-, 5 to 600-, 5 to 500-, 5 to 400-, 5 to 300-, 5 to 200-, 5 to 100-, 5 to 90-, 5 to 80-, 5 to 70-, 5 to 60-, 5 to 50-, 5 to 40-, 5 to 30-, 5 to 20-, 5 to 10-, 5 to 9-, 5 to 8-, 5 to 7-, 5 to 6-, 6 to 1000-, 6 to 900-, 6 to 800-, 6 to 700-, 6 to 600-, 6 to 500-, 6 to 400-, 6 to 300-, 6 to 200-, 6 to 100-, 6 to 90-, 6 to 80-, 6 to 70-, 6 to 60-, 6 to 50-, 6 to 40-, 6 to 30-, 6 to 20-, 6 to 10-, 6 to 9-, 6 to 8-, 6 to 7-, 7 to 1000-, 7 to 900-, 7 to 800-, 7 to 700-, 7 to 600-, 7 to 500-, 7 to 400-, 7 to 300-, 7 to 200-, 7 to 100-, 7 to 90-, 7 to 80-, 7 to 70-, 7 to 60-, 7 to 50-, 7 to 40-, 7 to 30-, 7 to 20-, 7 to 10-, 7 to 9-, 7 to 8-, 8 to 1000-, 8 to 900-, 8 to 800-, 8 to 700-, 8 to 600-, 8 to 500-, 8 to 400-, 8 to 300-, 8 to 200-, 8 to 100-, 8 to 90-, 8 to 80-, 8 to 70-, 8 to 60-, 8 to 50-, 8 to 40-, 8 to 30-, 8 to 20-, 8 to 10-, 8 to 9-, 9 to 1000-, 9 to 900-, 9 to 800-, 9 to 700-, 9 to 600-, 9 to 500-, 9 to 400-, 9 to 300-, 9 to 200-, 9 to 100-, 9 to 90-, 9 to 80-, 9 to 70-, 9 to 60-, 9 to 50-, 9 to 40-, 9 to 30-, 9 to 20-, 9 to 10-, 10 to 1000-, 10 to 900-, 10 to 800-, 10 to 700-, 10 to 600-, 10 to 500-, 10 to 400-, 10 to 300-, 10 to 200-, 10 to 100-, 10 to 90-, 10 to 80-, 10 to 70-, 10 to 60-, 10 to 50-, 10 to 40-, 10 to 30-, 10 to 20-, 20 to 1000-, 20 to 900-, 20 to 800-, 20 to 700-, 20 to 600-, 20 to 500-, 20 to 400-, 20 to 300-, 20 to 200-, 20 to 100-, 20 to 90-, 20 to 80-, 20 to 70-, 20 to 60-, 20 to 50-, 20 to 40-, 20 to 30-, 30 to 1000-, 30 to 900-, 30 to 800-, 30 to 700-, 30 to 600-, 30 to 500-, 30 to 400-, 30 to 300-, 30 to 200-, 30 to 100-, 30 to 90-, 30 to 80-, 30 to 70-, 30 to 60-, 30 to 50-, 30 to 40-, 40 to 1000-, 40 to 900-, 40 to 800-, 40 to 700-, 40 to 600-, 40 to 500-, 40 to 400-, 40 to 300-, 40 to 200-, 40 to 100-, 40 to 90-, 40 to 80-, 40 to 70-, 40 to 60-, 40 to 50-, 50 to 1000-, 50 to 900-, 50 to 800-, 50 to 700-, 50 to 600-, 50 to 500-, 50 to 400-, 50 to 300-, 50 to 200-, 50 to 100-, 50 to 90-, 50 to 80-, 50 to 70-, 50 to 60-, 60 to 1000-, 60 to 900-, 60 to 800-, 60 to 700-, 60 to 600-, 60 to 500-, 60 to 400-, 60 to 300-, 60 to 200-, 60 to 100-, 60 to 90-, 60 to 80-, 60 to 70-, 70 to 1000-, 70 to 900-, 70 to 800-, 70 to 700-, 70 to 600-, 70 to 500-, 70 to 400-, 70 to 300-, 70 to 200-, 70 to 100-, 70 to 90-, 70 to 80-, 80 to 1000-, 80 to 900-, 80 to 800-, 80 to 700-, 80 to 600-, 80 to 500-, 80 to 400-, 80 to 300-, 80 to 200-, 80 to 100-, 80 to 90-, 90 to 1000-, 90 to 900-, 90 to 800-, 90 to 700-, 90 to 600-, 90 to 500-, 90 to 400-, 90 to 300-, 90 to 200-, 90 to 100-, 100 to 1000-, 100 to 900-, 100 to 800-, 100 to 700-, 100 to 600-, 100 to 500-, 100 to 400-, 100 to 300-, 100 to 200-, 200 to 1000-, 200 to 900-, 200 to 800-, 200 to 700-, 200 to 600-, 200 to 500-, 200 to 400-, 200 to 300-, 300 to 1000-, 300 to 900-, 300 to 800-, 300 to 700-, 300 to 600-, 300 to 500-, 300 to 400-, 400 to 1000-, 400 to 900-, 400 to 800-, 400 to 700-, 400 to 600-, 400 to 500-, 500 to 1000-, 500 to 900-, 500 to 800-, 500 to 700-, 500 to 600-, 600 to 1000-, 600 to 900-, 600 to 800-, 600 to 700-, 700 to 1000-, 700 to 900-, 700 to 800-, 800 to 1000-, 800 to 900-, or 900 to 1000-fold reduction in the standard of care dose of a recombinant Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV protein vaccine. In some embodiments, the anti-antigenic polypeptide antibody titer produced in the subject is equivalent to an anti-antigenic polypeptide antibody titer produced in a control subject administered the standard of care dose of a recombinant or purified Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV protein vaccine or a live attenuated or inactivated Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV vaccine. In some embodiments, the effective amount is a dose equivalent to (or equivalent to an at least) 2-, 3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, 20-, 30-, 40-, 50-, 60-, 70-, 80-, 90-, 100-, 110-, 120-, 130-, 140-, 150-, 160-, 170-, 1280-, 190-, 200-, 210-, 220-, 230-, 240-, 250-, 260-, 270-, 280-, 290-, 300-, 310-, 320-, 330-, 340-, 350-, 360-, 370-, 380-, 390-, 400-, 410-, 420-, 430-, 440-, 450-, 4360-, 470-, 480-, 490-, 500-, 510-, 520-, 530-, 540-, 550-, 560-, 5760-, 580-, 590-, 600-, 610-, 620-, 630-, 640-, 650-, 660-, 670-, 680-, 690-, 700-, 710-, 720-, 730-, 740-, 750-, 760-, 770-, 780-, 790-, 800-, 810-, 820-, 830-, 840-, 850-, 860-, 870-, 880-, 890-, 900-, 910-, 920-, 930-, 940-, 950-, 960-, 970-, 980-, 990-, or 1000-fold reduction in the standard of care dose of a recombinant Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV protein vaccine. In some embodiments, an anti-antigenic polypeptide antibody titer produced in the subject is equivalent to an anti-antigenic polypeptide antibody titer produced in a control subject administered the standard of care dose of a recombinant or purified Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV protein vaccine or a live attenuated or inactivated Malaria (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale), JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV vaccine.

In some embodiments, the effective amount of a tropical disease RNA (e.g., mRNA) vaccine is a total dose of 50-1000 μg. In some embodiments, the effective amount of a tropical disease RNA (e.g., mRNA) vaccine is a total dose of 50-1000, 50-900, 50-800, 50-700, 50-600, 50-500, 50-400, 50-300, 50-200, 50-100, 50-90, 50-80, 50-70, 50-60, 60-1000, 60-900, 60-800, 60-700, 60-600, 60-500, 60-400, 60-300, 60-200, 60-100, 60-90, 60-80, 60-70, 70-1000, 70-900, 70-800, 70-700, 70-600, 70-500, 70-400, 70-300, 70-200, 70-100, 70-90, 70-80, 80-1000, 80-900, 80-800, 80-700, 80-600, 80-500, 80-400, 80-300, 80-200, 80-100, 80-90, 90-1000, 90-900, 90-800, 90-700, 90-600, 90-500, 90-400, 90-300, 90-200, 90-100, 100-1000, 100-900, 100-800, 100-700, 100-600, 100-500, 100-400, 100-300, 100-200, 200-1000, 200-900, 200-800, 200-700, 200-600, 200-500, 200-400, 200-300, 300-1000, 300-900, 300-800, 300-700, 300-600, 300-500, 300-400, 400-1000, 400-900, 400-800, 400-700, 400-600, 400-500, 500-1000, 500-900, 500-800, 500-700, 500-600, 600-1000, 600-900, 600-900, 600-700, 700-1000, 700-900, 700-800, 800-1000, 800-900, or 900-1000 μg. In some embodiments, the effective amount of a tropical disease RNA (e.g., mRNA) vaccine is a total dose of 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950 or 1000 μg. In some embodiments, the effective amount is a dose of 25-500 μg administered to the subject a total of two times. In some embodiments, the effective amount of a tropical disease RNA (e.g., mRNA) vaccine is a dose of 25-500, 25-400, 25-300, 25-200, 25-100, 25-50, 50-500, 50-400, 50-300, 50-200, 50-100, 100-500, 100-400, 100-300, 100-200, 150-500, 150-400, 150-300, 150-200, 200-500, 200-400, 200-300, 250-500, 250-400, 250-300, 300-500, 300-400, 350-500, 350-400, 400-500 or 450-500 μg administered to the subject a total of two times. In some embodiments, the effective amount of a tropical disease RNA (e.g., mRNA) vaccine is a total dose of 25, 50, 100, 150, 200, 250, 300, 350, 400, 450, or 500 μg administered to the subject a total of two times.

Examples of Additional Embodiments of the Disclosure

1. A tropical disease vaccine, comprising:

-   -   at least one messenger ribonucleic acid (mRNA) polynucleotide         having a 5′ terminal cap, an open reading frame encoding at         least one tropical disease antigenic polypeptide, and a 3′ polyA         tail.         2. The vaccine of paragraph 1, wherein the at least one tropical         disease antigenic polypeptide is selected from a Malaria         (e.g., P. falciparum, P. vivax, P. malariae and/or P. ovale)         antigenic polypeptide, a JEV antigenic polypeptide, a WNV         antigenic polypeptide, a EEEV antigenic polypeptide, a VEEV         antigenic polypeptide, a SINV antigenic polypeptide, a CHIKV         antigenic polypeptide, a DENV antigenic polypeptide, a ZIKV         antigenic polypeptide and a YFV antigenic polypeptide.         3. The vaccine of paragraph 1, wherein the at least one mRNA         polynucleotide is encoded by a sequence identified by any one of         SEQ ID NO: 1-6, 18, 19, 30-34, 48, 49, 55, 56, 65-80, 118-136,         223-239 or 376-388, or a fragment of a sequence identified by         any one of SEQ ID NO: 1-6, 18, 19, 30-34, 48, 49, 55, 56, 65-80,         118-136, 223-239 or 376-388.         4. The vaccine of paragraph 1, wherein the at least one mRNA         polynucleotide comprises a sequence identified by any one of SEQ         ID NO: 7-12, 20-21, 35-39, 50-51, 57-58, 81-96, 137-155,         240-256, or 389-401, or a fragment of a sequence identified by         any one of SEQ ID NO: 7-12, 20-21, 35-39, 50-51, 57-58, 81-96,         137-155, 240-256, or 389-401.         5. The vaccine of paragraph 1, wherein the at least one         antigenic polypeptide comprises a sequence identified by any one         of SEQ ID NO: 13-17, 22-29, 44-47, 52-54, 59-64, 97-117,         156-222, 469, 259-291 or 402-413, or a fragment of a sequence         identified by any one of SEQ ID NO: 13-17, 22-29, 44-47, 52-54,         59-64, 97-117, 156-222, 469, 259-291 or 402-413.         6. The vaccine of any one of paragraphs 1-5, wherein the 5′         terminal cap is or comprises 7mG(5′)ppp(5′)NlmpNp.         7. The vaccine of any one of paragraphs 1-6, wherein 100% of the         uracil in the open reading frame is modified to include         N1-methyl pseudouridine at the 5-position of the uracil.         8. The vaccine of any one of paragraphs 1-7, wherein the vaccine         is formulated in a lipid nanoparticle comprising: DLin-MC3-DMA;         cholesterol; 1,2-Distearoyl-sn-glycero-3-phosphocholine (DSPC);         and polyethylene glycol (PEG)₂₀₀₀-DMG.         9. The vaccine of paragraph 8, wherein the lipid nanoparticle         further comprises trisodium citrate buffer, sucrose and water.

This disclosure is not limited in its application to the details of construction and the arrangement of components set forth in the following description or illustrated in the drawings. The disclosure is capable of other embodiments and of being practiced or of being carried out in various ways. Also, the phraseology and terminology used herein is for the purpose of description and should not be regarded as limiting. The use of “including,” “comprising,” or “having,” “containing,” “involving,” and variations thereof herein, is meant to encompass the items listed thereafter and equivalents thereof as well as additional items.

EXAMPLES Example 1: Manufacture of Polynucleotides

According to the present disclosure, the manufacture of polynucleotides and/or parts or regions thereof may be accomplished utilizing the methods taught in International Publication WO2014/152027, entitled “Manufacturing Methods for Production of RNA Transcripts,” the contents of which is incorporated herein by reference in its entirety.

Purification methods may include those taught in International Publication WO2014/152030 and International Publication WO2014/152031, each of which is incorporated herein by reference in its entirety.

Detection and characterization methods of the polynucleotides may be performed as taught in International Publication WO2014/144039, which is incorporated herein by reference in its entirety.

Characterization of the polynucleotides of the disclosure may be accomplished using polynucleotide mapping, reverse transcriptase sequencing, charge distribution analysis, detection of RNA impurities, or any combination of two or more of the foregoing. “Characterizing” comprises determining the RNA transcript sequence, determining the purity of the RNA transcript, or determining the charge heterogeneity of the RNA transcript, for example. Such methods are taught in, for example, International Publication WO2014/144711 and International Publication WO2014/144767, the content of each of which is incorporated herein by reference in its entirety.

Example 2: Chimeric Polynucleotide Synthesis

According to the present disclosure, two regions or parts of a chimeric polynucleotide may be joined or ligated using triphosphate chemistry. A first region or part of 100 nucleotides or less is chemically synthesized with a 5′ monophosphate and terminal 3′desOH or blocked OH, for example. If the region is longer than 80 nucleotides, it may be synthesized as two strands for ligation.

If the first region or part is synthesized as a non-positionally modified region or part using in vitro transcription (IVT), conversion the 5′monophosphate with subsequent capping of the 3′ terminus may follow.

Monophosphate protecting groups may be selected from any of those known in the art.

The second region or part of the chimeric polynucleotide may be synthesized using either chemical synthesis or IVT methods. IVT methods may include an RNA polymerase that can utilize a primer with a modified cap. Alternatively, a cap of up to 130 nucleotides may be chemically synthesized and coupled to the IVT region or part.

For ligation methods, ligation with DNA T4 ligase, followed by treatment with DNase should readily avoid concatenation.

The entire chimeric polynucleotide need not be manufactured with a phosphate-sugar backbone. If one of the regions or parts encodes a polypeptide, then such region or part may comprise a phosphate-sugar backbone.

Ligation is then performed using any known click chemistry, orthoclick chemistry, solulink, or other bioconjugate chemistries known to those in the art.

Synthetic Route

The chimeric polynucleotide may be made using a series of starting segments. Such segments include:

(a) a capped and protected 5′ segment comprising a normal 3′OH (SEG. 1)

(b) a 5′ triphosphate segment, which may include the coding region of a polypeptide and a normal 3′OH (SEG. 2)

(c) a 5′ monophosphate segment for the 3′ end of the chimeric polynucleotide (e.g., the tail) comprising cordycepin or no 3′OH (SEG. 3)

After synthesis (chemical or IVT), segment 3 (SEG. 3) may be treated with cordycepin and then with pyrophosphatase to create the 5′ monophosphate.

Segment 2 (SEG. 2) may then be ligated to SEG. 3 using RNA ligase. The ligated polynucleotide is then purified and treated with pyrophosphatase to cleave the diphosphate. The treated SEG.2-SEG. 3 construct may then be purified and SEG. 1 is ligated to the 5′ terminus. A further purification step of the chimeric polynucleotide may be performed.

Where the chimeric polynucleotide encodes a polypeptide, the ligated or joined segments may be represented as: 5′UTR (SEG. 1), open reading frame or ORF (SEG. 2) and 3′UTR+PolyA (SEG. 3).

The yields of each step may be as much as 90-95%.

Example 3: PCR for cDNA Production

PCR procedures for the preparation of cDNA may be performed using 2×KAPA HIFI™ HotStart ReadyMix by Kapa Biosystems (Woburn, Mass.). This system includes 2× KAPA ReadyMix 12.5 μl; Forward Primer (10 μM) 0.75 μl; Reverse Primer (10 μM) 0.75 μl; Template cDNA 100 ng; and dH₂O diluted to 25.0 μl. The reaction conditions may be at 95° C. for 5 min. The reaction may be performed for 25 cycles of 98° C. for 20 sec, then 58° C. for 15 sec, then 72° C. for 45 sec, then 72° C. for 5 min, then 4° C. to termination.

The reaction may be cleaned up using Invitrogen's PURELINK™ PCR Micro Kit (Carlsbad, Calif.) per manufacturer's instructions (up to 5 μg). Larger reactions may require a cleanup using a product with a larger capacity. Following the cleanup, the cDNA may be quantified using the NANODROP™ and analyzed by agarose gel electrophoresis to confirm that the cDNA is the expected size. The cDNA may then be submitted for sequencing analysis before proceeding to the in vitro transcription reaction.

Example 4: In Vitro Transcription (IVT)

The in vitro transcription reaction generates RNA polynucleotides. Such polynucleotides may comprise a region or part of the polynucleotides of the disclosure, including chemically modified RNA (e.g., mRNA) polynucleotides. The chemically modified RNA polynucleotides can be uniformly modified polynucleotides. The in vitro transcription reaction utilizes a custom mix of nucleotide triphosphates (NTPs). The NTPs may comprise chemically modified NTPs, or a mix of natural and chemically modified NTPs, or natural NTPs.

A typical in vitro transcription reaction includes the following:

1) Template cDNA 1.0 μg 2) 10x transcription buffer 2.0 μl (400 mM Tris-HCl pH 8.0, 190 mM MgCl₂, 50 mM DTT, 10 mM Spermidine) 3) Custom NTPs (25 mM each) 0.2 μl 4) RNase Inhibitor 20 U 5) T7 RNA polymerase 3000 U 6) dH₂0 up to 20.0 μl. and 7) Incubation at 37° C. for 3 hr-5 hrs.

The crude IVT mix may be stored at 4° C. overnight for cleanup the next day. 1 U of RNase-free DNase may then be used to digest the original template. After 15 minutes of incubation at 37° C., the mRNA may be purified using Ambion's MEGACLEAR™ Kit (Austin, Tex.) following the manufacturer's instructions. This kit can purify up to 500 μg of RNA. Following the cleanup, the RNA polynucleotide may be quantified using the NANODROP™ and analyzed by agarose gel electrophoresis to confirm the RNA polynucleotide is the proper size and that no degradation of the RNA has occurred.

Example 5: Enzymatic Capping

Capping of a RNA polynucleotide is performed as follows where the mixture includes: IVT RNA 60 μg-180 μg and dH₂O up to 72 μl. The mixture is incubated at 65° C. for 5 minutes to denature RNA, and then is transferred immediately to ice.

The protocol then involves the mixing of 10× Capping Buffer (0.5 M Tris-HCl (pH 8.0), 60 mM KCl, 12.5 mM MgCl₂) (10.0 μl); 20 mM GTP (5.0 μl); 20 mM S-Adenosyl Methionine (2.5 μl); RNase Inhibitor (100 U); 2′-O-Methyltransferase (400U); Vaccinia capping enzyme (Guanylyl transferase) (40 U); dH₂O (Up to 28 μl); and incubation at 37° C. for 30 minutes for 60 μg RNA or up to 2 hours for 180 μg of RNA.

The RNA polynucleotide may then be purified using Ambion's MEGACLEAR™ Kit (Austin, Tex.) following the manufacturer's instructions. Following the cleanup, the RNA may be quantified using the NANODROP™ (ThermoFisher, Waltham, Mass.) and analyzed by agarose gel electrophoresis to confirm the RNA polynucleotide is the proper size and that no degradation of the RNA has occurred. The RNA polynucleotide product may also be sequenced by running a reverse-transcription-PCR to generate the cDNA for sequencing.

Example 6: PolyA Tailing Reaction

Without a poly-T in the cDNA, a poly-A tailing reaction must be performed before cleaning the final product. This is done by mixing capped IVT RNA (100 μl); RNase Inhibitor (20 U); 10× Tailing Buffer (0.5 M Tris-HCl (pH 8.0), 2.5 M NaCl, 100 mM MgCl₂) (12.0 μl); 20 mM ATP (6.0 μl); Poly-A Polymerase (20 U); dH₂O up to 123.5 μl and incubation at 37° C. for 30 min. If the poly-A tail is already in the transcript, then the tailing reaction may be skipped and proceed directly to cleanup with Ambion's MEGACLEAR™ kit (Austin, Tex.) (up to 500 μg). Poly-A Polymerase may be a recombinant enzyme expressed in yeast.

It should be understood that the processivity or integrity of the polyA tailing reaction may not always result in an exact size polyA tail. Hence, polyA tails of approximately between 40-200 nucleotides, e.g., about 40, 50, 60, 70, 80, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 150-165, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164 or 165 are within the scope of the present disclosure.

Example 7: Natural 5′ Caps and 5′ Cap Analogues

5′-capping of polynucleotides may be completed concomitantly during the in vitro-transcription reaction using the following chemical RNA cap analogs to generate the 5′-guanosine cap structure according to manufacturer protocols: 3′-O-Me-m7G(5′)ppp(5′) G [the ARCA cap]; G(5′)ppp(5′)A; G(5′)ppp(5′)G; m7G(5′)ppp(5′)A; m7G(5′)ppp(5′)G (New England BioLabs, Ipswich, Mass.). 5′-capping of modified RNA may be completed post-transcriptionally using a Vaccinia Virus Capping Enzyme to generate the “Cap 0” structure: m7G(5′)ppp(5′)G (New England BioLabs, Ipswich, Mass.). Cap 1 structure may be generated using both Vaccinia Virus Capping Enzyme and a 2′-O methyl-transferase to generate: m7G(5′)ppp(5′)G-2′-O-methyl. Cap 2 structure may be generated from the Cap 1 structure followed by the 2′-O-methylation of the 5′-antepenultimate nucleotide using a 2′-O methyl-transferase. Cap 3 structure may be generated from the Cap 2 structure followed by the 2′-O-methylation of the 5′-preantepenultimate nucleotide using a 2′-O methyl-transferase. Enzymes are preferably derived from a recombinant source.

When transfected into mammalian cells, the modified mRNAs have a stability of between 12-18 hours or more than 18 hours, e.g., 24, 36, 48, 60, 72 or greater than 72 hours.

Example 8: Capping Assays Protein Expression Assay

Polynucleotides (e.g., mRNA) encoding a polypeptide, containing any of the caps taught herein, can be transfected into cells at equal concentrations. The amount of protein secreted into the culture medium can be assayed by ELISA at 6, 12, 24 and/or 36 hours post-transfection. Synthetic polynucleotides that secrete higher levels of protein into the medium correspond to a synthetic polynucleotide with a higher translationally-competent cap structure.

Purity Analysis Synthesis

RNA (e.g., mRNA) polynucleotides encoding a polypeptide, containing any of the caps taught herein can be compared for purity using denaturing Agarose-Urea gel electrophoresis or HPLC analysis. RNA polynucleotides with a single, consolidated band by electrophoresis correspond to the higher purity product compared to polynucleotides with multiple bands or streaking bands. Chemically modified RNA polynucleotides with a single HPLC peak also correspond to a higher purity product. The capping reaction with a higher efficiency provides a more pure polynucleotide population.

Cytokine Analysis

RNA (e.g., mRNA) polynucleotides encoding a polypeptide, containing any of the caps taught herein can be transfected into cells at multiple concentrations. The amount of pro-inflammatory cytokines, such as TNF-alpha and IFN-beta, secreted into the culture medium can be assayed by ELISA at 6, 12, 24 and/or 36 hours post-transfection. RNA polynucleotides resulting in the secretion of higher levels of pro-inflammatory cytokines into the medium correspond to a polynucleotides containing an immune-activating cap structure.

Capping Reaction Efficiency

RNA (e.g., mRNA) polynucleotides encoding a polypeptide, containing any of the caps taught herein can be analyzed for capping reaction efficiency by LC-MS after nuclease treatment. Nuclease treatment of capped polynucleotides yield a mixture of free nucleotides and the capped 5′-5-triphosphate cap structure detectable by LC-MS. The amount of capped product on the LC-MS spectra can be expressed as a percent of total polynucleotide from the reaction and correspond to capping reaction efficiency. The cap structure with a higher capping reaction efficiency has a higher amount of capped product by LC-MS.

Example 9: Agarose Gel Electrophoresis of Modified RNA or RT PCR Products

Individual RNA polynucleotides (200-400 ng in a 20 μl volume) or reverse transcribed PCR products (200-400 ng) may be loaded into a well on a non-denaturing 1.2% Agarose E-Gel (Invitrogen, Carlsbad, Calif.) and run for 12-15 minutes, according to the manufacturer protocol.

Example 10: NANODROP™ Modified RNA Quantification and UV Spectral Data

Chemically modified RNA polynucleotides in TE buffer (1 μl) are used for NANODROP™ UV absorbance readings to quantitate the yield of each polynucleotide from an chemical synthesis or in vitro transcription reaction.

Example 11: Formulation of Modified mRNA Using Lipidoids

RNA (e.g., mRNA) polynucleotides may be formulated for in vitro experiments by mixing the polynucleotides with the lipidoid at a set ratio prior to addition to cells. In vivo formulation may require the addition of extra ingredients to facilitate circulation throughout the body. To test the ability of these lipidoids to form particles suitable for in vivo work, a standard formulation process used for siRNA-lipidoid formulations may be used as a starting point. After formation of the particle, polynucleotide is added and allowed to integrate with the complex. The encapsulation efficiency is determined using a standard dye exclusion assays.

Example 12: Immunogenicity Study

The instant study is designed to test the immunogenicity in mice of candidate Malaria vaccines comprising a mRNA polynucleotide encoding CS protein, LSA1, MSP1, AMA1, TRAP or a combination thereof obtained from Plasmodium.

Mice are immunized intramuscularly (IM), or intradermally (ID) with mRNA encoding CS protein, LSA1, MSP1, TRAP and AMA1. Up to three immunizations are given at 3-week intervals (i.e., at weeks 0, 3, and 6), and sera are collected after each immunization until weeks 33-51. Serum antibody titers against CS protein, LSA1, MSP1 and AMA1 are determined by ELISA. Responses against Plasmodium sporozoites, asexual blood-stage parasites, and gametocytes were determined by using an indirect immunofluorescence assay (IFA). T cell responses were analyzed by Elispot using splenocytes from immunized mice and stimulated with peptide pools from the relevant antigens.

Example 13: Plasmodium Non-Human Primate Challenge

The instant study is designed to test the efficacy in simians of candidate Malaria vaccines against a lethal challenge using a Malaria vaccine comprising mRNA encoding CS protein, LSA1, MSP1, AMA1, TRAP or a combination thereof obtained from Plasmodium. Simians are challenged with a lethal dose of Plasmodium.

Simians are immunized intramuscularly (IM) or intradermally (ID) at week 0, week 3 and week 6 with candidate Malaria vaccines.

Serum antibody titers against CS protein, LSA1, MSP1 and AMA1 are determined by ELISA. Responses against Plasmodium sporozoites, asexual blood-stage parasites, and gametocytes were determined by using an indirect immunofluorescence assay (IFA). T cell responses were analyzed by Elispot using PBMCs from immunized primates and stimulated with peptide pools from the relevant antigens.

In experiments where a lipid nanoparticle (LNP) formulation is used, the formulation may include a cationic lipid, non-cationic lipid, PEG lipid and structural lipid in the ratios 50:10:1.5:38.5. The cationic lipid may be DLin-KC2-DMA (50 mol %), the non-cationic lipid may be DSPC (10 mol %), the PEG lipid may be PEG-DOMG (1.5 mol %) and the structural lipid may be cholesterol (38.5 mol %), for example.

Example 14: Plasmodium Human Challenge

The instant study is designed to test the efficacy in human subjects of candidate Malaria vaccines against an attenuated challenge (Controlled Human Malaria Infection (CHMI) Study) using a Malaria vaccine comprising mRNA encoding CS protein, LSA1, MSP1, AMA1, TRAP or a combination thereof obtained from Plasmodium. Subjects are challenged with an attenuated (non-lethal) dose of Plasmodium.

Subjects are immunized intramuscularly (IM) or intradermally (ID) at week 0 and week 3 with candidate Malaria vaccines. Serum is tested for microneutralization (see Example 16). The subjects are then challenged with an attenuated dose of Plasmodium on week 7 via IV, IM or ID. Endpoint is day 13 post infection. Body temperature and weight are assessed and recorded daily.

In experiments where a lipid nanoparticle (LNP) formulation is used, the formulation may include a cationic lipid, non-cationic lipid, PEG lipid and structural lipid in the ratios 50:10:1.5:38.5. The cationic lipid may be DLin-KC2-DMA (50 mol %), the non-cationic lipid may be DSPC (10 mol %), the PEG lipid may be PEG-DOMG (1.5 mol %) and the structural lipid may be cholesterol (38.5 mol %), for example.

Example 15: Microneutralization Assay

Nine serial 2-fold dilutions (1:50-1:12,800) of simian or human serum are made in 50 μl virus growth medium (VGM) with trypsin in 96 well microtiter plates. Fifty microliters of Plasmodium are added to the serum dilutions and allowed to incubate for 60 minutes at room temperature (RT). Positive control wells of Plasmodium without sera and negative control wells without Plasmodium or sera are included in triplicate on each plate. While the serum-Plasmodium mixtures incubate, a single cell suspension of cells is prepared by trypsinizing (Gibco 0.5% bovine pancrease trypsin in EDTA) a confluent monolayer, and suspended cells are transferred to a 50 ml centrifuge tube, topped with sterile PBS and gently mixed. The cells are then pelleted at 200 g for 5 minutes, supernatant aspirated and cells resuspended in PBS. This procedure is repeated once and the cells are resuspended at a concentration of 3×10⁵/ml in VGM with porcine trypsin. Then, 100 μl of cells are added to the serum-virus mixtures and the plates incubated at 35° C. in CO₂ for 5 days. The plates are fixed with 80% acetone in phosphate buffered saline (PBS) for 15 minutes at RT, air dried and then blocked for 30 minutes containing PBS with 0.5% gelatin and 2% FCS. An antibody to CS protein, LSA1, MSP1, AMA1 or TRAP is diluted in PBS with 0.5% gelatin/2% FCS/0.5% Tween 20 and incubated at RT for 2 hours. Wells are washed and horse radish peroxidase conjugated goat anti-mouse IgG added, followed by another 2 hour incubation. After washing, 0-phenylenediamine dihydrochloride is added and the neutralization titer is defined as the titer of serum that reduced color development by 50% compared to the positive control wells.

Example 16: JEV Immunogenicity Study

This study was designed to test the immunogenicity of JEV prME mRNA vaccines in Balb/c mice. Mice were 6-8 weeks old.

Mice were immunized intramuscularly at three different doses (10 μg, 2 μg and 0.5 μg). All mice were given two doses of the vaccine, one at day 0 and another at day 28. Serum was collected at days 0 and 56, and a plaque reduction neutralization test was used to quantify neutralizing antibody titer. The concentration of serum to reduce the number of plaques in the assay by 50%, compared to the serum free virus, denoted as PRNT50 was used as a measure of neutralizing antibodies and level of protection against virus.

Results of this study is shown in FIG. 1. A PRNT50 titer of greater than 1:10 is considered protective. JEV mRNA vaccine at 10 μg doses results in a very high titer, indicative of a high potency vaccine.

Example 17: Immunogenicity Cross-Neutralization Study

The instant study is designed to test the immunogenicity and cross-neutralization in mice of candidate combination vaccines comprising a mRNA polynucleotide encoding antigenic polypeptides (e.g., envelope proteins) obtained from Plasmodium, JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV.

Mice are immunized intravenously (IV), intramuscularly (IM), or intradermally (ID) with candidate combination vaccines. A total of four immunizations are given at 3-week intervals (at weeks 0, 3, 6, and 9), and sera are collected after each immunization until weeks 33-51. Serum antibody titers against envelope proteins are determined by ELISA. Sera collected from each mouse during weeks 10-16 are pooled, and total IgGs are purified by using ammonium sulfate (Sigma) precipitation followed by DEAE (Pierce) batch purification. Following dialysis against PBS, the purified antibodies are used for immunoelectron microscopy, antibody-affinity testing, and an in vitro protection assay.

Example 18: Immunogenicity Studies for Combination RNA Vaccine

BALB/C mice are immunized with mRNA encoded Plasmodium, JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptides, for example, as shown in Table 7 below and according to the following dosing/bleeding schedule: prime dose on day 0, boost dose on day 28, bleeding on days 0, 28, 42 and 56.

The mice are administered a combination vaccine, combining two or more of the Plasmodium, JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV antigenic polypeptides, such that all possible combinations are tested. Animals are challenged at day 56 with a second dose of any one of the antigenic polypeptides included in the original dose.

An efficacy study using mRNA encoded West Nile prMEs and Japanese Encephalitis prMEs antigens is also performed according to the following schedule:

Non-human primates are also immunized with mRNA encoded antigen using similar schedules shown in Tables 7 and 8. The animals are tested for immunogenicity to Plasmodium, JEV, WNV, EEEV, VEEV, SINV, CHIKV, DENV, ZIKV and/or YFV and combinations thereof.

Example 19: YFV Immunogenicity Studies

The instant study is designed to test the immunogenicity in Balb/c mice of candidate yellow fever virus (YFV) vaccines comprising a mRNA polynucleotide encoding YFV prME. Four groups of Balb/c mice (n=5) are immunized intramuscularly (IM) with 10 μg (n=2) or 2 μg (n=2) of the candidate vaccine. One group of mice is administered PBS intramuscularly as a control. All mice are administered an initial dose of vaccine (Groups 1-4) or PBS (Group 5) on Day 0, and then the mice in Groups 1 and 3 are administered a boost dose on Day 21, while the mice in Group 5 are administered PBS on Day 21. All mice are bled on Day 41. See Table 1. Anti-Yellow fever neutralization IgG titer is determined on Day −1, Day 28 and Day 41.

Example 20: YFV Rodent Challenge

The instant study is designed to test the efficacy in AG129 mice of candidate yellow fever virus (YFV) vaccines against a lethal challenge using a YFV vaccine comprising mRNA encoding YFV prME. Four groups of AG129 mice (n=8) are immunized intramuscularly (IM) with 10 μg (n=2) or 2 μg (n=2) of the candidate vaccine. One group of mice is administered PBS intramuscularly as a control. All mice are administered an initial dose of vaccine (Groups 1-4) or PBS (Group 5) on Day 0, and then the mice in Groups 1 and 3 are administered a boost dose on Day 21, while the mice in Group 5 are administered PBS on Day 21. All mice are challenged with a lethal dose of YFV in Day 42. All mice are then monitored for survival and weight loss. Anti-Yellow fever neutralization IgG titer is determined on Day −1, Day 28 and Day 41, and viral load is determined 5 days post challenge.

Example 21: Expression of ZIKV prME Protein in Mammalian Cells Using ZIKV mRNA Vaccine Construct

The Zika virus (ZIKV) prME mRNA vaccine construct were tested in mammalian cells (239T cells) for the expression of ZIKV prME protein. 293T cells were plated in 24-well plates and were transfected with 2 μg of ZIKV prME mRNA using a Lipofectamine transfection reagent. The cells were incubated for the expression of the ZIKV prME proteins before they were lysed in an immunoprecipitation buffer containing protease inhibitor cocktails. Reducing agent was not added to the lysis buffer to ensure that the cellular proteins were in a non-reduced state. Cell lysates were centrifuged at 8,000×g for 20 mins to collect lysed cell precipitate. The cell precipitates were then stained with anti ZIKV human serum and goat anti-human Alexa Fluor 647. Fluorescence was detected as an indication of prME expression.

The expression of ZIKV prME protein was also detected by fluorescence-activated cell sorting (FACS) using a flow cytometer. 293F cells (2×10⁶ cells/ml, 30 ml) were transfected with 120 μg PEI, 1 ml of 150 mM NaCl, and 60 μg prME mRNA. Transfected cells were incubated for 48 hours at 37° C. in a shaker at 130 rpm and under 5% CO₂. The cells were then washed with PBS buffer containing 2% FBS and fixed in a fixation buffer (PBS buffer containing formalin) for 20 minutes at room temperature. The fixed cells were permeabilized in a permeabilization buffer (PBS+1% Triton X100+1 μl of Golgi plug/ml of cells). The permeabilized cells were then stained with anti-ZIKV human serum (1:20 dilution) and goat anti-human Alexa Fluor 647 secondary antibody, before they were sorted on a flow cytometer. As shown in FIG. 2, FIG. 3A and FIG. 3B, cells transfected with prME mRNA and stained with the anti-ZIKA human serum shifted to higher fluorescent intensity, indicating that prME expressed from the ZIKV mRNA vaccine constructs in the transfected cells.

Example 22: Expression, Purification and Characterization of ZIKV VLPs

Zika virus (ZIKV) virus-like particles (VLPs) were made in HeLa cells and in HEK293T cells and purified via PEG precipitation or ultracentrifugation, respectively. Cells were cultured in culture media. Prior to transfection, cells were passaged twice in virus growth media plus 10% fetal bovine serum (FBS) to media adaptation.

Cells were seeded the day before transfection into T-175 flask. 100 μg of prME-encoding mRNA was transfected using 100 μg pf lipofectamine as per manufacturer's protocol. 6 hours post transfection, monolayers were washed twice with 1×PBS and 20 mL of virus growth media was added. Supernatant was collected 24-48 hours post transfection by centrifugation at 2000×g for 10 mins and 0.22 μm filtration.

For VLP purification via PEG precipitation, VLP's were concentrated using Biovision PEG precipitation kit as per manufacturer's protocol. In brief, supernatant with VLP's was mixed with PEG8000 and incubated at 4° C. for 16 hours. After incubation, mixture was centrifuged at 3000×g for 30 mins. Pellet containing concentrated VLP's was collected and suspended into PBS. VLP's were further buffer exchanged into PBS (1:500) using amicon ultra 100 MWCO filter. Purified samples were negative stained to show the presence of assembled VLP particles.

Expression of prME from the vaccine mRNA constructs was demonstrated to result in the production of virus like particles (VLPs) that are expected to present to the immune system as identical to Zika virus particles. Negative stain electron micrographs of supernatants from HeLa cells transfected with mRNA encoding Zika prME showed that the virus-like particles (VLPs), purified by PEG precipitation, have highly uniform size (˜35-40 nm) and morphology. The bumpy appearance of the VLP surface appears to reflect mostly immature morphology due to expression from HeLa cells, which have very low expression of furin, a host protease that is required for maturation the viral envelope. Upon maturation, these VLPs will have an exterior structure essentially identical to wild type viral particles, thus eliciting a broad immune response to future Zika virus exposure.

For VLP purification via ultracentrifugation, 293T cells were transfected with Zika prME mRNA as described herein. Supernatant was collected 24 hours after changing the media as described herein (30 hours post transfection). VLPs were concentrated using Biovision PEG virus precipitation kit into 500 μL volume. VLPs were further purified using a 10-50% sucrose gradient. Sample layer was seen between 20-30% sucrose layers and collected. VLPs were buffered exchanged into PBS by 1:1000 dilution using a 100 MWCO amicon ultra filter. VLPs were concentrated after PEG precipitation, and ultracentrifuge-purified VLPs were analyzed for purity on a reducing SDS-PAGE gel (FIG. 4).

Example 23: ZIKV mRNA Vaccine Immunogenicity Studies

The instant study was designed to test the immunogenicity in Balb/c mice of candidate ZIKV vaccines comprising a mRNA polynucleotide encoding ZIKV prME. Four groups of Balb/c mice (n=5) were immunized intramuscularly (IM) with 10 μg (n=2) or 2 μg (n=2) of the candidate vaccine. One group of mice was administered PBS intramuscularly as a control. All mice were administered an initial dose of vaccine (Groups 1-4) or PBS (Group 5) on Day 0, and then the mice in Groups 1 and 3 were administered a boost dose on Day 21, while the mice in Group 5 were administered PBS on Day 21. All mice were bled on Day 41. See Table 29. Anti-Zika neutralization IgG titer was determined on Day −1, Day 28 and Day 41 (FIG. 5).

Day 42 neutralizing titers reached EC50s of 427 for 2 μg and 690 for 10 μg. The control serum in this experiment was from naturally infected immunocompromised mice (Ifnar1−/−, derived from B/6 lineage) in which high viral loads would be achieved.

Example 24: ZIKV Rodent Challenge

The instant study was designed to test the efficacy in AG129 mice of candidate ZIKV vaccines against a lethal challenge using a ZIKV vaccine comprising mRNA encoding ZIKV prME. Four groups of AG129 mice (n=8) were immunized intramuscularly (IM) with 10 μg (n=2) or 2 μg (n=2) of the candidate vaccine. One group of mice was administered PBS intramuscularly as a control. All mice were administered an initial dose of vaccine (Groups 1-4) or PBS (Group 5) on Day 0, and then the mice in Groups 1 and 3 were administered a boost dose on Day 21, while the mice in Group 5 were administered PBS on Day 21. All mice were challenged with a lethal dose of ZIKV in Day 42. All mice were then monitored for survival and weight loss. Anti-Zika neutralization IgG titer was determined on Day −1, Day 28 and Day 41, and viral load was determined 5 days post challenge. The 10 μg dose provided 100% protection, even with a single dose, and the 2 μg dose provided 60% protection with a single dose and 90% protection with prime-boost doses (see FIGS. 7A and 7B).

In experiments where a lipid nanoparticle (LNP) formulation is used, the formulation may include a cationic lipid, non-cationic lipid, PEG lipid and structural lipid in the ratios 50:10:1.5:38.5. The cationic lipid may be DLin-KC2-DMA or DLin-MC3-DMA (50 mol %), the non-cationic lipid may be DSPC (10 mol %), the PEG lipid is PEG-DOMG or PEG-DMG (1.5 mol %) and the structural lipid may be cholesterol (38.5 mol %), for example.

Example 25: Exemplary Dengue Sequences

An exemplary Dengue virus (DENY) peptide epitope may include two or more epitopes. The epitopes can be of the same sequence or different sequence and can be all T-cell epitopes, all B-cell epitopes or a combination of both. Furthermore, various end units for enhancing MHC processing of the peptides are possible.

The following sequences represent exemplary DENY peptide epitopes identified using a database screen (the sequences correspond to SEQ ID NO: 357-360):

DenV1 1

80 DenV2 1

80 DenV3 1

80 DenV4 1

80 DenV1 81

160 DenV2 81

160 DenV3 81

158 DenV4 81

160 DenV1 161

240 DenV2 161

240 DenV3 159

238 DenV4 161

240 DenV1 241

320 DenV2 241

320 DenV3 239

318 DenV4 241

320 DenV1 321

400 DenV2 321

394 DenV3 319

392 DenV4 321

400

Nucleic acid and amino acid sequences for each of DENV-1, DENV-2, DENV-3, and DENV-4 are shown in Tables 28 and 29, respectively.

Example 26: Dengue Virus RNA Vaccine Immunogenicity in Mice

This study provides a preliminary analysis of the immunogenicity of a nucleic acid mRNA vaccine using a Dengue virus (DENV) serotype 2 antigen in BALB/c mice. The study utilizes 44 groups of 10 BALB/c female (5) and male (5) mice (440 total, 6-8 weeks of age at study initiation, see Table 10 for design summary). In this study, construct numbers used are referenced and found in Table 28.

Mice were vaccinated on weeks 0 and 3 via intramuscular (IM) or intradermal (ID) routes. One group remained unvaccinated and one was administered 10⁵ plaque-forming units (PFU) live DENV2, D2Y98P isolate via intravenous (IV) injection as a positive control. Serum was collected from each mouse on weeks 1, 3, and 5; bleeds on weeks 1 and 3 were in-life samples (tail vein or submandibular bleeds) and week 5 will be a terminal (intracardiac) bleed. Individual serum samples were stored at −80° C. until analysis by neutralization or microneutralization assay. Pooled samples from each group at the week 5 time points were tested by Western blot for reactivity with viral lysate.

Signal was detected in groups 5, 15, 39, and 44 (live virus control) by a band that appeared between 50 and 60 kDa in the Western blot data. The data suggests that a mRNA vaccine to a single dengue viral antigen can produce antibody in preliminary studies.

In order to provide a Dengue vaccine having enhanced immunogenicity, RNA vaccines for concatemeric antigens were designed and tested according to the invention. These vaccines, which have significantly enhanced activity, in comparison to the single protein antigens described herein, are described below.

Example 27: In Silico Prediction of T Cell Epitopes for RNA Vaccine Design

Several peptide epitopes from Dengue virus were generated and tested for antigenic activity. The peptide epitopes are designed to maximize MHC presentation. In general the process of MHC class I presentation is quite inefficient, with only 1 peptide of 10,000 degraded molecules actually being presented. Additionally the priming of CD8 T cell with APCs having insufficient densities of surface peptide/MHC class I complexes results in weak responders exhibiting impaired cytokine secretion and a decrease memory pool. Thus, the process of designing highly effective peptide epitopes is important to the immunogenicity of the ultimate vaccine.

In silico prediction of desirable peptide epitopes was performed using Immune Epitope Database. Using this database several immunogenic Dengue T cell epitopes showing strong homology across all 4 Dengue serotypes were predicted. Examples of these epitopes are shown in FIGS. 8A-8C and 9A-9C.

Example 28: Prediction of DENV T Cell Epitopes for RNA Vaccine Design

The design of optimized vaccination systems to prevent or treat conditions that have failed to respond to more traditional treatments or early vaccination strategies relies on the identification of the antigens or epitopes that play a role in these conditions and which the immune system can effectively target. T cell epitopes (e.g., MHC peptide binding) for the various alleles shown in Table 32 were determined using Rapid Epitope Discovery System (Prolmmune REVEAL & ProVE®—see Tables 33-40 for peptides). This system is used to identify those candidate epitopes that actually cause relevant immune responses from the numerous other potential candidates identified using algorithms to predict MHC-peptide binding. The REVEAL binding assay determines the ability of each candidate peptide to bind to one or more MHC I class alleles and stabilize the MHC-peptide complex. The assay identifies the most likely immunogenic peptides in a protein sequence by comparing the binding to that of a high affinity T cell epitope and detecting the presence or absence of the native conformation of the MHC-peptide complex. The epitope peptides are further tested using the assays described herein to confirm their immunogenic activity.

Example 29: Activity Testing for Predicted Peptide Epitopes

Exemplary peptide epitopes selected using the methods described above were further characterized. These peptide epitopes were confirmed to have activity using in vitro HLA binding assays (human lymphocyte binding assays). Peptides (9 aa peptides from the dengue antigen) were screened for their ability to bind to HLA. The analysis of the homology, affinity, frequency and design of these peptides is shown in FIGS. 8A-8C and 9A-9C.

Example 30: In Vivo Analysis of Mimectopes of Predicted Human Epitopes RNA Vaccines Methods

IFNγ ELISpot. Mouse IFNγ ELISpot assays were performed using IFNγ coated Millipore IP Opaque plates according to the manufacturer's mouse IFNγ ELISPOT guidelines. Briefly, the plates were blocked using complete RPMI (R10) and incubated for 30 minutes prior to plating cells. Peptides (284-292, 408-419 or 540-548) were diluted to 5 different concentrations for stimulation at 5, −6, −7, −8, or −9 from an original stock concentration of 10 mM⁽⁻²⁾. Mouse splenocytes (200,000-250,000 cells) were plated in appropriate wells with peptide, PMA+lonomycin or R₁₀ media alone. Cells were stimulated in a total volume of 125 μL per well. Plates were then incubated at 37° C., 5% CO₂ for 18-24 hrs. Plates were developed following the manufacturer's instructions. Plates were counted and quality controlled using the automated ELISPOT reader CTL ImmunoSpot/FluoroSpot.

Intracellular Cytokine Staining (ICS).

Intracellular Cytokine Staining (ICS). For intracellular cytokine staining, individual splenocytes, were resuspended at a concentration of 1.5×10⁶ cells per mL. Peptides (284-292, 408-419 or 540-548) were made into 5 dilutions from a stock concentration of 10 mM⁽⁻²⁾. The final concentrations of each peptide were −5, −6, −7, −8, or −9 in their respective wells. Cells were stimulated in a final volume of 200 μL within a 96 well culture plate. After the addition of Golgi plug (0.2 μL per well), cells were incubated at 37° C., 5% CO₂ for 5 hours. Following stimulation, cells were surface stained, fixed, washed and put at 4° C. overnight. Intracellular staining was performed the following day, resulting in full panel of Live/Dead (Invitrogen), αCD3, αCD4, αCD8, αCD45, αCCR7, αCD44, αCD25, αIL-2, αTNγ, and αTNFα (BD Biosciences). Cells were acquired in a 96-U bottom plate using BD LSR Fortessa HTS (BD Biosciences).

Results

The exemplary peptide epitopes selected using the methods described herein were used to produce tests mouse mimectopes of the predicted human epitopes. These mimectopes were analyzed for in vivo activity using restimulation assays during the acute phase of Dengue infection (Day 7). The methods were performed on dengue-infected IFNαβ/γ-receptor-deficient mice (AG129). Seven days post infection splenocytes were harvested and subjected to an ELISPOT assay to quantify secretion of cytokines by T cells (CD8) as described above. Briefly, the isolated splenocytes were stimulated with the test peptides and tested for T cell activation. If the peptide is an appropriate antigen, some cells would be present antigen during infection and would be capable of stimulating T cells. The methods for analyzing the T cell activation were performed as follows:

-   -   T cells (at a known concentration) were incubated with a         specific antigen in a cell culture well     -   the activated T cells were transferred to ELISPOT plates         (precoated with anti-cytokine antibody)     -   the cells were incubated such that cytokines could be secreted     -   the cells were washed off the plate and enzyme coupled secondary         Ig was added     -   the plates were washed and substrate was added     -   positive spots were scored under microscope.

The data is shown in FIGS. 10 and 11. FIGS. 10 and 11 are graphs depicting the results of an ELISPOT assay of dengue-specific peptides measuring IFN-γ (spots per million splenocytes).

A schematic of an assay on a BLT Mouse Model (Bone Marrow/Liver/Thymus) is shown in FIG. 12. The results of a histogram analysis of human CD8 T cells stimulated with peptide epitope is also shown in FIG. 12.

The following two sequences were used as controls:

(V5)8-cathb: (SEQ ID NO: 361) Kozak Start GKPIPNPLLGLDST-GFLG-GKPIPNPLLGLDST- GFLG-GKPIPNPLLGLDST-GFLG-GKPIPNPLLGLDST-GFLG- GKPIPNPLLGLDST-GFLG-GKPIPNPLLGLDST-GFLG- GKPIPNPLLGLDST-GFLG-GKPIPNPLLGLDST Stop (v5)8-cathb + MHCi: (SEQ ID NO: 362) Kozak Start GKPIPNPLLGLDST-GFLG-GKPIPNPLLGLDST- GFLG-GKPIPNPLLGLDST-GFLG-GKPIPNPLLGLDST-GFLG- GKPIPNPLLGLDST-GFLG-GKPIPNPLLGLDST-GFLG- GKPIPNPLLGLDST-GFLG-GKPIPNPLLGLDST Stop Some results are shown in Table 41.

Example 31: AG129 Mouse Challenge of Mimectopes of Predicted Human Epitopes from DENV2

A study is performed on AG129 mouse using a cocktail of 2 peptide epitopes. The immunogenicity of the peptide epitopes is determined in AG129 mice against challenge with a lethal dose of mouse-adapted DENV 2 strain D2Y98P. AG129 mice, which lack IFN α/β and γ receptor signaling, injected intradermally in the footpad with 10⁴ PFU of DENV do not survive past day 5 post-injection. AG129 mice are vaccinated via intramuscular (IM) injection with either 2 μg or 10 μg of a cocktail of 2 peptide epitopes. The vaccines are given to AG129 mice with a prime and a boost (day 0 and day 28). The positive control group is vaccinated with heat-inactivated DENV 2. Phosphate-buffered saline (PBS) is used as a negative control. On day 56, mice are challenged with mouse-adapted DENV 2 and monitored for 10 days for weight loss, morbidity, and mortality. Mice that display severe illness, defined as >30% weight loss, a health score of 6 or above, extreme lethargy, and/or paralysis are euthanized.

Example 32: “Humanized” DENV Peptides Mouse Immunogenicity Study

A study analyzing immunogenicity of the peptide epitopes on humanized mice is performed. A single-dose cocktail (30 μg) containing 3 different peptide epitopes are delivered by IM route of immunization with prime and boost (day 0, day 28). A T cell (ELISPOT and ICS) characterization may be performed on Day 7, Day 28, and Day 56.

Example 33: Testing of Non-Human Primate (NHP) Mimectopes of Predicted DENV Human Epitopes

Non-human primate (NHP) mimectopes to the human epitopes may also be developed and tested for activity in NHP assays. The NHP mimectopes are designed based on the human antigen sequence. These mimectopes may be analyzed for in vivo activity in an NHP model using, for instance, restimulation assays. Once the NHPs have been infected, immune cells may be isolated and tested for sensitivity of activation by the particular mimectopes.

Example 34: Targeting of DENV Concatemeric Constructs Using Cytoplasmic Domain of MHC I

MHC-1_V5 concatemer constructs were developed and transfected in HeLa cells. Triple immunofluorescence using Mitotracker Red (mitochondria), anti-V5, and anti-MHC-1 antibodies plus DAPI was performed. MHC-1_V5 concatemer transfection in HeLa cells shows V5-MHC1 colocalization. MHC-1 V5 concatemer transfection also shows V5 has homogeneous cytoplasmic distribution and preferentially colocalizes with MHC1 and not with Mitotracker. These data demonstrate that the V5 concatemer with the cytoplasmic domain from MHC class I co-localizes with MHC class I expression, while the V5 concatemer without this sequence is only found in the cytoplasm following transfection in HeLa cells.

Example 35: In Vivo Analysis of DENV Concatemeric mRNA Epitope Construct

The Dengue concatemers used in this study consist of 8 repeats of the peptide TALGATEI (SEQ ID NO: 363), a mouse CD8 T cell epitope found in the DENV2 envelope. The peptide repeats were linked via cathepsin B cleavage sites and modified with the various sequences as follows:

-   -   (1) TALGATEI (SEQ ID NO: 363) peptide concatemer with no         modification     -   (2) TALGATEI (SEQ ID NO: 363) peptide concatemer with IgKappa         signal peptide     -   (3) TALGATEI (SEQ ID NO: 363) peptide concatemer with PEST         sequence     -   (4) TALGATEI (SEQ ID NO: 363) peptide concatemer with IgKappa         signal peptide and PEST sequence     -   (5) TALGATEI (SEQ ID NO: 363) peptide concatemer with MHC class         I cytoplasmic domain     -   (6) TALGATEI (SEQ ID NO: 363) peptide concatemer with IgKappa         signal peptide and MHC class I cytoplasmic domain     -   (7) Heat-inactivated DENV2 (D2Y98P)     -   (8) No immunization

The immunogenicity of the peptide concatemeric candidate vaccines was determined in AG129 mice against challenge with a lethal dose of DENV strain D2Y98P. AG129 mice, which lack IFN α/β and γ receptor signaling, injected intradermally in the footpad with 10⁴ PFU of DENV do not survive past day 5 post-injection. (In this study, the mice died due to a problem with the heat-attenuation). The tested vaccines included constructs (1)-(8) disclosed above. AG129 mice were vaccinated via intramuscular (IM) injection with either 2 μg or 10 μg of the candidate vaccine. The vaccines were given to AG129 mice as a prime and a boost (second dose provided 28 days after the first dose). The positive control group was vaccinated with heat-inactivated DENV2. Phosphate-buffered saline (PBS) was used as a negative control.

On day 56, mice were challenged with mouse-adapted DENV2 and monitored for 10 days for weight loss, morbidity, and mortality. Mice that displayed severe illness, defined as >30% weight loss, a health score of 6 or above, extreme lethargy, and/or paralysis were euthanized. Notably, mice “vaccinated” with heat-inactivated DENV (positive control group) became morbid and died (they were not included in the challenge portion of the study).

In addition, individual serum samples were collected prior to challenge on day 54 and PBMCs were isolated and frozen for subsequent testing.

The AG129 mice PBMCs were thawed and stimulated with TALGATEI (SEQ ID NO: 363) peptide for 5 hours in a standard intracellular cytokine assay. For intracellular cytokine staining, PBMCs were thawed and suspended in media. The TALGATEI (SEQ ID NO: 363) peptide was administered to stimulate the cells. After the addition of Golgi plug, cells were incubated at 37° C., 5% CO₂ for 5 hours. Following stimulation, cells were surface stained, fixed, washed and put at 4° C. overnight. Intracellular staining was performed the following day and assayed via ELISPOT assay to quantify secretion of cytokines by T cells (CD8) as described above to determine T cell activation. If the peptide were an appropriate antigen, some cells would be present antigen during infection and would be capable of stimulating T cells. The results are shown in FIGS. 13A and 13B, which demonstrate that each of the peptides (1)-(6) stimulate T cell activation.

Example 36: Surface-Expressed DENV2 prME Antigens

The DENV2 prME polypeptide antigen sequences provided in Tables 28 and 29 were tested to confirm that the DENV prME protein antigen is translated, properly folded and expressed on the surface of cells. For the polypeptide sequences, the bolded sequence is Dengue signal sequence, the underlined sequence is DENV2 precursor membrane sequence, and the unmarked sequence is DENV2 envelope sequence. The sequences encoding the polypeptides are codon-optimized. HeLa cells were transfected with DNA encoding the prMEs from nine different DENV2 isolates. After 24 hours, surface expression of the prME was detected using three different antibodies followed by goat-anti-human AF700 secondary antibody and subjecting the cells to FACS analyses. Each of the three antibodies is broadly neutralizing DENV2 prME antibodies that have in vivo efficacy against Dengue virus. D88 binds to DIII of Envelope protein for all 4 DENV serotypes (US20150225474). 2D22 binds to DIII of Envelope protein for DENV 2 serotype. 5J7 binds to 3 domains of Envelope protein for DENV 3 serotype. FIG. 14B shows that the D88 and 2D22 antibodies recognize two of the DENV2 prME antigens. These results show that the two DENV2 prME antigens identified as Thailand/01 68/1979 and Peru/IQT29 13/1996 are expressed at the cell surface in a conformationally correct form and are excellent vaccine candidates (FIG. 14A). FIG. 14B shows a repeat of staining in triplicate and in two different cell lines (HeLa and 293T). These results confirm proper conformation of expressed DENV2 prME antigens (in particular, the prME antigens from Thailand/01 68/1979 and Peru/IQT29 13/1996) and also evidence at least non-inferior and even superior DENV2 antigenicity as compared to Dengvaxia (CYD-TDV), a live attenuated tetravalent chimeric vaccine. Antigen expressed from the mRNA encoding DENV 2 prME from Peru/IQT2913/1996 shows the best binding to 2 different DENV2 antibodies in 293T cells and in HeLa cells (D88—binds all 4 serotypes 2D22—binds DENV 2). This construct has a single amino acid difference from the DENV 2 Envelope III Domain immunodeterminant region (see bold, underline in SEQ ID NO: 273, DENV 2 prME (Peru/IQT2913/1996) in Table 29).

Example 37: OVA Multitope In Vitro Screening Assay Kinetic Analysis

Antigen surface presentation is an inefficient process in the antigen presenting cells (APC). Peptides generated from proteasome degradation of the antigens are presented with low efficiency (only 1 peptide of 10000 degraded molecules is actually presented). Thus, priming of CD8 T cells with APCs provides insufficient densities of surface peptide/MHC I complexes, resulting in weak responders exhibiting impaired cytokine secretion and decreased memory pool. To improve DENV mRNA vaccines encoding concatemeric DENV antigens, an in vitro assay was designed to test the linkers used to connect peptide repeats, the number of peptide repeats, and sequences known to enhance antigen presentation.

mRNA constructs encoding one or more OVA epitopes were configured with different linker sequences, protease cleavage sites, and antigen presentation enhancer sequences. Their respective sequences were as shown in Table 43. To perform the assay, 200 ng of each MC3-formulated mRNA construct was transfected into JAWSII cells in a 24-well plate. Cells were isolated at 6, 24, and 48 hours post transfection and stained with fluorescently-labeled Anti-Mouse OVA257-264 (SIINFEKL (SEQ ID NO: 364)) peptide bound to H-2 Kb. Staining was analyzed on a LSRFortessa flow cytometer. Samples were run in triplicate. The Mean Fluorescent Intensity (MFI) for each mRNA construct was measured and shown in FIG. 15. Constructs 2, 3, 7, 9, and 10 showed enhanced surface presentation of the OVA epitope, indicating that the configurations of these constructs may be used for DENV mRNA vaccine. Construct 5 comprises a single OVA peptide and a KDEL sequence that is known to prevent the secretion of a protein. Construct 5 showed little surface antigen presentation because the secretion of the peptide was inhibited.

Example 38: Antibody Binding to DENV-1, 2, 3, and 4 prME Epitopes

DENV mRNA vaccines encoding concatemeric antigen epitopes were tested for binding to antibodies known to recognize one or more DENV serotypes. To test antibody binding to the epitopes, 200 ng of DENV mRNA vaccines encoding different Dengue prME epitopes were transfected into HeLa cells in 24-well plates using the TransitIT-mRNA Transfection Kit (Mirus Bio). The DENV mRNA vaccine constructs are shown in Table 28. Transfections were done in triplicate. After 24 hours, surface expression was detected using four different antibodies (10 μg/mL) followed by either goat-anti-human or anti-mouse AF700 secondary antibody (1/500). Signal generated from antibody binding are shown as Mean Fluorescent Intensity (MFI) (FIG. 16). Antibody D88 is known to recognize all 4 serotypes and bound to all antigen epitopes encoded by the DENV mRNA vaccine constructs tested. Antibody 2D22 is known to recognize only DENV 2 and preferentially bound to construct 21, which encodes DENV 2 antigen epitopes. Antibody 2D22 also showed weak binding to epitopes of other DENV serotypes. Antibody 5J7 is known to recognize only DENV 3 and only bound to antigen epitopes encoded by constructs 13, 19, and 20, which encode DENV 3 antigen epitopes. Antibody 1-11 is known to bind strongly to DENV 1 and 2, to bind weakly to DENV 3 and to bind little DENV 4. Antibody 1-11 bound to DENV 1, 2, and 3, and binding to DENV 3 antigen epitopes was stronger than binding to DENV 1 or 2 (FIG. 16).

Example 39: DENV prME Challenge Study in Cynomolgus (Cyno) Monkey Model

Shown in Table 45 is the design of DENV prME challenge study in cynomolgus (cyno) money. Indicated DENV mRNA vaccine encoding prME antigen epitopes, or vaccines thereof, are used to immunize cyno. The vaccines are formulated in lipid nanoparticles (e.g., MC3 formulation) and administered to the cyno monkeys intramuscularly on day 0, 21, and 42. Dosages of the vaccines are 250 μg or 5 μg per immunization. In experiments where a combination of different DENV mRNA vaccines are used, 250 μg or 5 μg of each mRNA vaccine is used. FLAG-tagged H10N8 flu vaccine is used as control at a dosage of 250 μg per immunization. Naïve cyno monkeys without immunization are also used as control. Cyno monkey sera are collected on days 20, 41, 62, and 92 post initial immunization and used for serotype-specific neutralization assays.

Immunized cyno monkeys are challenged on day 63 post initial immunization with indicated DENV viruses. Cyno monkey sera are collected on days 62 (pre-challenge), 63-66, 68, 70, 72, 76, and 92 (end of life) to determine serum viral load.

Example 40: Dengue 2 prME Challenge Study in AG129 Mice

The instant study was designed to evaluate the efficacy of four DENV mRNA vaccine constructs (constructs 21-24 in Table 44) in AG129 mice challenge assays. The schedule of the challenge study is shown in FIG. 17A. The DENV mRNA vaccines were formulated in lipid nanoparticles (e.g., MC3 formulation) and administered to the AG129 mice intramuscularly on days 0 and 21. Dosages of the vaccines were 2 μg or 10 μg per immunization. Heat inactivated D2Y98P strain was used as a negative control to vaccinate the mice. Naïve AG129 mice without immunization were also used as control.

Immunized AG129 mice were challenged on day 42 post initial immunization with Dengue D2Y98P virus (s.c., 1e5 PFU per mouse). AG129 mice sera were collected on days 20 and 41 post initial immunization and used for serotype-specific neutralization assays. Mice immunized with any of the four DENV mRNA vaccine constructs survived, while the control mice died. These data demonstrate that, after lethal challenge, there was 100% protection provided by each mRNA vaccine construct, regardless of dose. The weights and health of the mice were monitored and the results were plotted in FIGS. 17C-17D.

Mice sera collected from mice immunized with 2 μg of the DENV mRNA vaccines were able to neutralize several DENV 2 strains and variations in the neutralization ability between the tested mRNA vaccines and between different DENV 2 strains were observed (FIG. 18).

Example 41: DENV prME Challenge Study in AG129 Mouse Model

Shown in Table 46 is the design of a DENV prME challenge study in AG129 mice, including the mRNA constructs tested, the vaccination schedule, the dosage, the challenge strains, and the serum collection schedule.

Indicated DENV mRNA vaccines encoding prME antigen epitopes, or vaccines thereof, were used to immunize AG129 mice. The vaccines were formulated in lipid nanoparticles (e.g., MC3 formulation) and administered to the mice intramuscularly on days 0 and 21. Dosages of the vaccines were 2 μg or 10 μg per immunization. In experiments where a combination of different DENV mRNA vaccines was used, 2 μg of each mRNA vaccine was used. Naïve AG129 mice without immunization were used as control. AG129 mice sera were collected on days 20 and 41 post initial immunization and used for serotype-specific neutralization assays.

Immunized AG129 mice were challenged on day 42 post initial immunization with Dengue D2Y98P virus (s.c., 1e5 PFU per mouse). The weights and health of the mice were monitored for 14 days post infection and the results were plotted in FIGS. 19A-19I.

Example 42: Virus-Like Particles

The antigens produced from the DENV prME mRNA vaccines of the present disclosure, when expressed, are able to assemble into virus-like particles (VLPs). The instant study was designed to evaluate the immunogenicity of the VLPs by negative stain electron microscope imaging. DENV mRNA vaccine constructs 21-24 were expressed and VLPs were assembled an isolated. The VLPs were visualized under negative stain electron microscopy. Construct 23 is the vaccine construct used by Sanofi in its DENV vaccines. Constructs 21, 22, and 24 produced more uniform VLPs, suggesting that these VLPs may be more superior in their immunogenicity than the VLPs produced from construct 23.

Example 43: Exemplary Nucleic Acids Encoding CHIKV RNA Polynucleotides for Use in a RNA Vaccine

Exemplary sequences that can be used to encode CHIKV E1, E2, E1-E2, and C-E3-E2-6K-E1 RNA polynucleotides for use in the CHIKV RNA vaccine are given in Table 47.

Example 44: Protocol to Determine Efficacy of mRNA-Encoded Chikungunya Antigen Candidates Against CHIKV

Chikungunya virus (CHIKV) has a polycistronic genome and different antigens, based on the Chikungunya structural protein, are possible. There are membrane-bound and secreted forms of E1 and E2, as well as the full length polyprotein antigen, which retains the protein's native conformation. Additionally, the different CHIKV genotypes can also yield different antigens.

The efficacy of CHIKV candidate vaccines in AG129 mice against challenge with a lethal dose of CHIKV strain 181/25 was investigated. A129 mice, which lack IFN α/β receptor signaling, injected intradermally in the footpad with 10⁴ PFU of CHIKV 181/25 virus have a 100% survival rate post-injection. In contrast, AG129 mice, which lack IFN α/β and γ receptor signaling, injected intradermally in the footpad with 10⁴ PFU of CHIKV 181/25 virus do not survive past day 5 post-injection. The tested vaccines included: MC3-LNP formulated mRNA encoded CHIKV-E1, MC3-LNP formulated mRNA encoded CHIKV-E2, and MC3-LNP formulated mRNA encoded CHIKV-E1/E2/E3/C. Fifteen groups of five AG129 mice were vaccinated via intradermal (ID) or intramuscular (IM) injection with either 2 μg or 10 μg of the candidate vaccine. The vaccines were given to AG129 mice as single or two doses (second dose provided 28 days after the first dose). The positive control group was vaccinated via intranasal instillation (20 μL volume) with heat-inactivated CHIKV. Phosphate-buffered saline (PBS) was used as a negative control.

On day 56, mice were challenged with 1×10⁴ PFU of CHIKV via ID injection in a 50 μL volume and monitored for 10 days for weight loss, morbidity, and mortality. Mice that displayed severe illness, defined as >30% weight loss, a health score of 6 or above, extreme lethargy, and/or paralysis were euthanized. Notably, mice “vaccinated” with heat-inactivated CHIKV (positive control group) became morbid and were euthanized following the second dose of HI-CHIKV (they were not included in the challenge portion of the study).

In addition, individual samples were tested for reactivity in a semi-quantitative ELISA for mouse IgG against either Chikungunya-specific E1 (groups 1-4), Chikungunya-specific E2 (groups 5-8), or Chikungunya-specific E1 and E2 proteins (groups 9-15).

The health status is scored as indicated in Table 51.

Example 45: Efficacy of Chikungunya E1 Antigen mRNA Vaccine Candidate

AG129 mice (n=5 per group) were vaccinated with 2 μg or 10 μg of MC-3-LNP formulated mRNA encoding CHIKV E1. The AG129 mice were vaccinated on either Day 0 or Days 0 and 28 via IM or ID delivery. On Day 56 following final vaccination all mice were challenged with a lethal dose of CHIKV. The survival curve, percent weight loss, and health status of the mice vaccinated with 2 μg CHIKV E1 mRNA are shown in FIGS. 21A-21C. The survival results are tabulated in Table 52. The survival curve, percent weight loss, and health status of the mice vaccinated with 10 μg CHIKV E1 mRNA are shown in FIGS. 24A-24C. The survival results are tabulated in Table 53.

As shown in Table 52, the 2 μg dose of CHIKV E1 mRNA vaccine gave no protection post-CHIKV infection challenge when administered via IM or ID with either a single dose or two doses. Likewise, the single dose of 10 μg CHIKV E1 vaccine provided little to no protection when administered via IM or ID. However, as indicated in Table 53, the 10 μg dose of CHIKV E1 mRNA vaccine provided 60% protection post-CHIKV challenge when administered via IM using two doses and provided 80% protection post-CHIKV challenge when administered via ID using two doses.

In all experiments, the negative control mice had a ˜0% survival rate, as did the positive control mice (heat-inactivated CHIKV), which died before CHIKV challenge. Some mice died during the vaccination period.

Example 46: Efficacy of Chikungunya E2 Antigen mRNA Vaccine Candidate

AG129 mice (n=5 per group) were vaccinated with 2 μg or 10 μg of MC-3-LNP formulated mRNA encoding CHIKV E2. The mice were vaccinated on either Day 0 or Days 0 and 28 via IM or ID delivery. On Day 56 following final vaccination all mice were challenged with a lethal dose of CHIKV. The survival curve, percent weight loss, and health status of the mice vaccinated with 2 μg CHIKV E2 mRNA are shown in FIGS. 22A-22C. The survival results are tabulated in Table 54 below. The survival curve, percent weight loss, and health status of the mice vaccinated with 10 μg CHIKV E2 mRNA are shown in FIGS. 25A-25C. The survival results are tabulated in Table 55.

As shown in Table 54, the 2 μg dose of CHIKV E2 mRNA vaccine gave no protection post-CHIKV infection challenge when administered via IM or ID in a single dose. However, when provided in two doses, the 2 μg dose of CHIKV E2 mRNA vaccine provided 80% protection when administered via IM and 100% protection when administered via ID post-CHIKV challenge. As indicated in Table 55, the 10 μg dose of CHIKV E2 mRNA mouse provided no protection post-CHIKV challenge when administered via IM or ID in a single dose. However, administration of CHIKV E2 mRNA via IM or ID using two doses provided 100% protection post-CHIKV challenge.

In all experiments, the negative control mice had a ˜0% survival rate, as did the positive control mice (heat-inactivated CHIKV) which died prior to CHIKV challenge. Some mice died during the vaccination period.

Example 47: Efficacy of Chikungunya C-E3-E2-6K-E1 Antigen mRNA Vaccine Candidate

AG129 mice (n=5 per group) were vaccinated with 2 μg or 10 μg of MC-3-LNP formulated mRNA encoding CHIKV C-E3-E2-6K-E1 mRNA (SEQ ID NO: 388/401). The AG129 mice were vaccinated on either Day 0 or Days 0 and 28 via IM or ID delivery. On Day 56 following final vaccination all mice were challenged with a lethal dose of CHIKV. The survival curve, percent weight loss, and health status of the mice vaccinated with 2 μg CHIKV C-E3-E2-6K-E1 mRNA are shown in FIGS. 23A-23C. The survival results are tabulated in Table 56. The survival curve, percent weight loss, and health status of the mice vaccinated with 10 μg CHIKV C-E3-E2-6K-E1/E2/E3/C mRNA are shown in FIGS. 26A-26C. The survival results are tabulated in Table 57.

As shown in Table 56, the 2 μg dose of C-E3-E2-6K-E1 mRNA vaccine provided 100% protection post-CHIKV challenge when administered via IM in a single dose and provided 80% protection post-CHIKV challenge when administered via ID in a single dose. The 2 μg dose of C-E3-E2-6K-E1 mRNA vaccine provided 100% protection post-CHIKV challenge when administered via IM or ID in two doses. As shown in Table 57, the 10 μg dose of C-E3-E2-6K-E1 mRNA vaccine provided 100% protection post-CHIKV infection challenge when administered via IM or ID in either a single dose or in two doses.

In all experiments, the negative control mice had a ˜0% survival rate, as did the positive control mice (heat-inactivated CHIKV) which died prior to CHIKV challenge. Some mice died during the vaccination period.

Example 48: Summary of Survival Data Using Chikungunya Antigen mRNA Vaccine Candidates CHIKV E1, CHIKV E2, and CHIKV C-E3-E2-6K-E1

Table 58 shows the survival data of the mice vaccinated with the CHIKV mRNA antigens used in the studies reported in Examples 45-47.

Example 49: In Vitro Transfection of mRNA-Encoded Chikungunya Virus Envelope Protein

The in vitro transfection of mRNA encoding Notch and a PBS control were performed in 150 k HeLa cells/well transfected with 1 μg mRNA+2 μL LF2000/well in a 24 well plate. Lysate containing proteins expressed from the CHIKV envelope mRNAs transfected in HeLa cells were collected 16 hours post-transfection and then detected by Western blotting with a V5 tag-HRP antibody. The successful detection of a CHIKV envelope protein is shown in FIG. 20.

Example 50: Detection of Immunity (Mouse IgG) Against Either Chikungunya-Specific E1, Chikungunya-Specific E2, or Chikungunya-Specific E1 and E2 Proteins

Serum samples from mice vaccinated with the CHIKV E1, E2, or E1-E2-E3-C vaccine described in Examples 45-47 were tested using a semi-quantitative ELISA for the detection of mouse IgG against either Chikungunya-specific E1, Chikungunya-specific E2, or Chikungunya-specific E1 and E2 proteins.

Fifteen groups of five mice were vaccinated via intradermal (ID) or intramuscular (IM) injection with either 2 μg or 10 μg of the candidate vaccine. The vaccines were given to AG129 mice as single or two doses (second dose provided 28 days after the first dose). On day 56, mice were challenged with 1×104 PFU of CHIKV via ID injection in 50 μL volume and monitored for 10 days for weight loss, morbidity, and mortality. Mice were bled on day 7 and day 28 post-vaccination via the peri-orbital sinus (retro-orbital bleed). In addition, mice surviving the CHIKV challenge were bled 10 days post-challenge.

The individual samples were tested for reactivity in a semi-quantitative ELISA for mouse IgG against either Chikungunya-specific E1, Chikungunya-specific E2, or Chikungunya-specific E1 and E2 proteins. The results are shown in FIGS. 34-36.

The data depicting the results of the ELISA assay to identify the amount of antibodies produced in AG129 mice in response to vaccination with mRNA encoding secreted CHIKV E1 structural protein, secreted CHIKV E2 structural protein, or CHIKV full structural polyprotein C-E3-E2-6k-E1 at a dose of 10 μg or 2 μg at 28 days post immunization is shown in FIGS. 34-35. The 10 μg of mRNA encoding CHIKV polyprotein produced significant levels of antibody in both studies. The data depicting a comparison of ELISA titers from the data of FIG. 34 to survival in the data of FIG. 35 left panel is shown in FIG. 36. As shown in the survival results, the animals vaccinated with either dose (single or double administration) of mRNA encoding CHIKV polyprotein had 100% survival rates.

Example 51: Efficacy of Chikungunya Polyprotein (C-E3-E2-6K-E1) mRNA Vaccine Candidate

AG129 mice (n=5 per group) were vaccinated with either 10 μg, 2 μg or 0.4 μg of MC-3-LNP formulated mRNA encoded CHIKV polyprotein (C-E3-E2-6K-E1) (SEQ ID NO: 388/401). The mice were vaccinated on either Day 0 or Days 0 and 28 via IM delivery. In one study, all mice were challenged on day 56 with a lethal dose of CHIKV following final vaccination. In another study, all mice were challenged on day 84 with a lethal dose of CHIKV following final vaccination. The survival curve, percent weight loss, and health status of the mice vaccinated with 10 μg, 2 μg or 0.4 μg mRNA were determined as described previously in Examples 45-47. The survival rates, neutralizing antibodies and binding antibodies were assessed. Neutralizing antibodies were also identified against three different strains of CHIKV.

The survival rates of the mice vaccinated with mRNA encoding CHIKV C-E3-E2-6k-E1 is shown in FIG. 37. The data depicts vaccination at a dose of 10 μg (left panels), 2 μg (middle panels) or 0.4 μg (right panels) at 56 days (top panels) or 112 days (bottom panels) post immunization. These data demonstrate that a single 2 μg dose of the mRNA vaccine afforded 100% protection for at least 112 days (16 weeks). Following the study out further, the data demonstrated that a single 2 μg dose of the mRNA vaccine afforded 100% protection for at least 140 days (20 weeks.)

The neutralizing antibody and binding antibody produced in treated mice is shown in FIGS. 38 and 39, respectively. As can be seen in FIGS. 38 and 39, the levels of neutralizing antibody were dependent or dose and regimen with the highest titers evident with 10 μg dosed twice (days 0 and 28). Plaque reduction neutralization tests (PRNT50 and PRNT80) were used to quantify the titer of neutralizing antibody for the virus. Antigen-binding Ab was determined by ELISA. The corresponding correlation between binding Ab and neutralizing antibodies is shown in the bottom panels of FIG. 39. Following the study out to 16 weeks showed that the highest E1 titers were achieved when 10 μg mRNA vaccine was dosed twice.

The data depicting neutralizing antibodies against three different strains of CHIKV is shown in FIG. 40. The neutralizing antibodies were tested against three different strains of CHIKV, African—Senegal (left panel), La Reunion (middle panel) and CDC CAR (right panel). FIG. 40 shows that the polyprotein-encoding mRNA vaccine elicited broadly neutralizing antibodies against the three strains tested. Sera were further tested against Chik S27 strain (Chikungunya virus (strain S27-African prototype). The data depicting neutralizing antibodies against CHIKV S27 strain is shown in FIG. 41. These data collectively show that the polyprotein encoding mRNA vaccine elicited broadly neutralizing antibodies against all four strains tested. The vaccine induced neutralizing antibodies against multiple strains of Chikungunya. The prime and boost with the 10 μg dose produced the most robust neutralizing antibody response followed by the single dose with 10 μg.

Example 52: Transfection of mRNA Encoded CHIKV Structural Proteins

In vitro transfection of mRNA encoding CHIKV structural proteins and PBS control were performed in 400 k HeLa cells transfected with 1.25 ug mRNA lipoplexed with 5 ul LF2000/well in 6 well plate. Protein detection in HeLa cell lysate 16 h post transfection was measured. Lysates which contain proteins expressed from the CHIKV mRNAs transfected in HeLa were collected 16 h post transfection. Proteins were detected by WB with anti-Flag or and V5 antibody.

The mRNA encoded CHIKV structural proteins and protein production in the HeLa cell lysate 16 h post transfection was detected.

Example 53: Exemplary CHIKV Polypeptides

The amino acids presented in the Table 48 are exemplary CHIKV antigenic polypeptides. To the extent that any exemplary antigenic peptide described herein includes a flag tag or V5, or a polynucleotide encodes a flag tag or V5, the skilled artisan understands that such flag tag or V5 is excluded from the antigenic polynucleotide in a vaccine formulation. Thus, any of the polynucleotides encoding proteins described herein are encompassed within the compositions of the invention without the flag tag or V5 sequence.

Example 54: Efficacy of CHIKV mRNA Vaccine X Against CHIKV in AG129 Mice Study Design

Chikungunya virus (CHIKV) 181/25 strain is an attenuated vaccine strain that was developed by the US Army via multiple plaque-to-plaque passages of the 15561 Southeast Asian human isolate (Levitt et al.). It is well tolerated in humans and is highly immunogenic. It produces small plaques and has decreased virulence in infant mice and nonhuman primates. When the attenuated vim s is administered to immunodeficient AG129 mice (lacking the IFN-α/β and γ receptors) the mice succumb to a lethal disease within 3-4 days with ruffled fur and weight loss (Partidos, et al. 2011 Vaccine).

This instant study was designed to evaluate the efficacy of CHIKV candidate vaccines as described herein in AG129 mice (Table 59). The study included 14 groups of female 6-8 week old AG129 mice (Table 59). Groups 1-4, 7-8, and 10-15 were vaccinated with CHIKV vaccine X via the intramuscular (IM; 0.05 mL) route on Day 0 and select groups received an additional boost on Day 28. Control Groups 9 and 16 received vehicle (PBS) only on Days 0 and 28 via IM route (0.05 mL). Regardless of vaccination schedule, Groups 1-4 and 7-9 were challenged on Day 56 while Groups 10-16 were challenged on Day 112 using the CHIKV 181/25 strain (stock titer 3.97×10⁷ PFU/mL, challenge dose 1×10⁴ PFU/mouse). For virus challenge, all mice received a lethal dose (1×10⁴ PFU) of Chikungunya (CHIK) strain 181/25 via intradermal (ID) route (0.050 mL via footpad). All mice were monitored for 10 days post infection for weight loss, morbidity, and mortality. Each mice was assigned a heath score based on Table 51. Mice displaying severe illness as determined by >30% weight loss, a health score of higher than 5, extreme lethargy, and/or paralysis were euthanized with a study endpoint of day 10 post virus challenge. Test bleeds via retro-orbital (RO) collection were performed on mice from all groups on Days −3, 28, and 56. Mice from Groups 10-16 were also bled on Days 84 & 112. Mice that survived challenge were also terminally bled on Day 10 post challenge. Serum samples from mice (Days −3, 28, 56, 84, 112 and surviving mice) were kept frozen (−80° C.) and stored until they were tested for reactivity in a semi quantitative ELISA for mouse IgG against either E1, E2 or CHIKV lysate.

Experimental Procedure Intramuscular (IM) Injection of Mice

1. Restrain the animal either manually, chemically, or with a restraint device.

2. Insert the needle into the muscle. Pull back slightly on the plunger of the syringe to check proper needle placement. If blood is aspirated, redirect the needle and recheck placement again.

3. Inject appropriate dose and withdraw needle. Do not exceed maximum volume. If the required volume exceeds the maximum volume allowed, multiple sites may be used with each receiving no more than the maximum volume.

4. The injection site may be massaged gently to disperse the injected material.

Intradermal (ID) Injections of Mice

1. Restrain the animal either manually, chemically, or with a restraint device.

2. Carefully clip the hair from the intended injection site. This procedure can be done upon animals arriving or the day before any procedures or treatments are required.

3. Lumbar area is the most common site for ID injections in all species, but other areas can be used as well.

4. Pinch or stretch the skin between your fingers (or tweezers) to isolate the injection site.

5. With the beveled edge facing up, insert the needle just under the surface between the layers of skin. Inject the appropriate dose and withdraw needle. A small bleb will form when an ID injection is given properly.

6. If the required volume exceeds the maximum volume allowed, multiple sites may be used with each receiving no more than the maximum volume.

Retro-Orbital Bleeding in Mice

1. Place the mice in the anesthesia chamber and open oxygen line and set to 2.5% purge. Start flow of anesthesia at 5% isoflurane.

2. Once the animal becomes sedate, turn anesthesia to 2.5%-3% isoflurane and continue to expose the animal to the anesthesia. Monitor the animal to avoid breathing becoming slow.

3. Remove the small rodent from anesthesia chamber and place on its back while restraining with left hand and scruff the back of the animal's neck, so it is easy to restrain and manipulate while performing the procedure with the right hand.

4. With a small motion movement, place the capillary tube in the corner of the animal's eye close to the nostril, and rotate or spin the Hematocrit glass pipette until blood start flowing out. Collect the appropriate amount of blood needed into the appropriate labeled vial.

5. Monitor the animal after retro-orbital bleeding is done for at least 10-15 seconds to ensure hemostasis.

6. Place the animal back to its original cage and monitor for any other problems or issues caused while manipulating animal due to the procedure.

Observation of Mice

1. Mice were observed through 10 days post infection (11 days total, 0-10 days post infection).

2. Mice were weighed daily on an Ohause scale and the weights are recorded.

3. Survival and health of each mouse were evaluated once time a day using a scoring system of 1-7 described in Table 51.

Infection

On either Day 56 (Groups 1-4, 7-9) or Day 112 (Groups 10-16) groups of 5 female 6-8 week old AG129 mice were infected via intradermal injection with 1×10⁴ PFU/mouse of the 181/25 strain of Chikungunya diluted in PBS. The total inoculation volume was 0.05 mL administered in the rear footpad of each animal. Mice were anesthetized lightly using 2-5% v/v of isoflurane at ˜2.5 L/min of O2 (VetEquip IMPAC6) immediately prior to infection.

Dose Administration

In this study mice were administered 0.04 μg, 2 μg, or 10 μg of various formulations of the CHIKV vaccine X or vehicle alone (PBS) on either Day 0 or on Days 0 and 28 via the intramuscular route (0.05 mL). The material was pre-formulated and diluted in PBS by IBT prior to dosing.

Results

Mice were immunized once (Day 0) or twice (Days 0 & 28) with either 0.04 μg, 2 μg, or 10 μg of Chikungunya mRNA vaccine X and were challenged with CHIKV strain 181/25 on either Day 56 (Groups 1-4, 7-9) or on Day 112 (Groups 10-16). Mice were monitored for a total of 10 days post infection for health and weight changes. Mice that received either 2 μg or 10 μg of the CHIKV mRNA vaccine X either once (Day 0) or twice (Days 0 and 28) were fully protected (100%) regardless of whether the mice were challenged 56 days or 112 days after the initial vaccination (FIGS. 27A-27B, Table 44). Mice receiving 0.04 μg of the CHIKV mRNA vaccine were not protected at all from lethal CHIKV infection. This efficacy data is supported by the health scores observed in the vaccinated mice in that the protected mice displayed little to no adverse health effects of a CHIKV infection (FIGS. 29A-29B). Weight loss is not a strong indicator of disease progression in the CHIKV AG129 mouse model (FIGS. 28A-28B).

Mice immunized with the CHIKV mRNA vaccine X showed increased antibody titers against CHIKV E1, E2 and CHIKV lysate as compared to the vehicle only (PBS) treated groups. Serum binding against the virus lysate yielded the highest antibody titers for all vaccinated groups (FIGS. 30A-30C, 31A-31C, 32A-32C, 33A-33C). Overall, the antibody titers were dose dependent with the highest titers observed in serum from mice vaccinated with 10 μg of CHIKV mRNA vaccine X while the lowest titers were observed in serum from mice vaccinated with 0.04 μg of the CHIKV mRNA vaccine X. Similarly, higher titers were observed in serum from mice vaccinated twice (Days 0 and 28) as compared to serum from mice vaccinated only once (Day 0). Serum obtained on Day 112 post initial vaccination still yielded increased antibody titers in mice that received either 10 μg or 2 μg of CHIKV mRNA vaccine X (FIGS. 32A-32C).

Serum from mice groups 10-16, 112 days post immunization were also tested in a Plaque Reduction Neutralization Test (PRNT). Serum from each mice was diluted from 1/20 to 1/40960 and assessed for its ability to reduce CHIKV plaque formation. The results were shown in Table 64.

Example 55: Immunogenicity of Chikungunya Polyprotein (C-E3-E2-6K-E1) mRNA Vaccine Candidate in Rats

Sprague Dawley rats (n=5) were vaccinated with 20 μg of MC-3-LNP formulated mRNA 30 encoded CHIKV polyprotein (C-E3-E2-6K-E1) (SEQ ID NO: 388/401). The rats were vaccinated on either Day 0 or Days 0 and 14 or Days 0, 14 and 28 via IM delivery. Sera were collected on days −3, 14, 28 and 42 for ELISA testing. FIG. 42 demonstrated that there was at least a two log increase in antibody titer against CHIKV lysate post 3rd vaccination with the mRNA vaccine in normal rats.

Example 56: Evaluation of T Cell Activation of Chikungunya P 5 Polyprotein (C-E3-E2-6K-E1) mRNA Vaccine Candidate

C57BL/6 mice (n=6 experimental group; n=3 control group) were vaccinated with 10 μg of MC-3-LNP formulated mRNA encoded CHIKV polyprotein (C-E3-E2-6K-E1) (SEQ ID NO: 388/401). The mice were vaccinated on either Day 0 or Days 0 and 28 (boost) via IM delivery. Sera was collected on days 3, 28 and 42 for ELISA testing. Animals were sacrificed on day 42 and spleens were harvested for immunological evaluation of T cells. Splenic cells were isolated and analyzed by FACS. Briefly, spleens were removed, cells isolated, and stimulated in vitro with immunogenic peptides found within either C, E1, or E2 region of CHIKV that are known to be CD8 epitopes in B6 mice. The readout for this assay was cytokine secretion (IFN-gamma and TNF-alpha), which reveals whether the vaccine induced antigen-specific T cell responses. No CD8 T cell responses were detected using the E2 or C peptide (baseline levels of IFN-gamma and TNF-alpha), whereas there was a response to the E1-corresponding peptide (average of about 0.4% IFN-gamma and 0.1% TNF). The peptides were used to stimulate T cells used in the study were E1=HSMTNAVTI (SEQ ID NO: 414), E2=IILYYYELY (SEQ ID NO: 415), and C=ACLVGDKVM (SEQ ID NO: 416).

FIG. 43 shows that the polyprotein-encoding CHIKV polyprotein vaccine elicited high antibody titers against the CHIKV glycoproteins. FIGS. 44 and 45A-45B show T cell activation by E1 peptide.

Example 57: Proof-of-Concept of Immunogenicity in Non-Human Primates

The mRNA vaccine was tested in Cynomolgus monkey subjects (n=3 per experimental group, n=3 negative control). Subjects were given an intramuscular (IM) immunization of 25 μg or 75 μg of the vaccine on day 0 (prime), day 28 (boost), and day 56 (boost). The negative control group was administered 75 μg of non-translated irrelevant mRNA (NTIX). The readout for this experiment was serum antibody titers (binding and neutralizing) and a CHIKV-specific T cell response.

As shown in FIG. 46, the vaccine induced a robust antibody response. A response was detected after the priming dose, and then increased with the boost, and increased slightly more following the third immunization. Both the 25 μg and 75 μg vaccine groups were immunogenic, and there was a small dose response. Neutralizing titers were a few fold lower than those seen in mice, but were still robust.

FIG. 47 shows a robust CD4 response in response to the vaccine. Day 35 T cells, measured one week after the second immunization, were assayed. The peptide pool consisted of 15mers overlapping by 11. The response was measured through peptide stimulation, followed by intracellular cytokine staining and flow cytometry. A CHIKV-specific CD4 T cell response was detected, mainly in IL-2 and TNFα. There was a minimal CD8 response as well.

Each of the sequences described herein encompasses a chemically modified sequence or an unmodified sequence (no modified nucleotides), which includes no nucleotide modifications.

TABLE 1 Plasmodium Nucleic Acid Sequences SEQ ID Description Sequences NO: CSP_Full_length ATGATGAGAAAATTAGCTATTTTATCTGTTTCTTCCTTTTTATTTG  1 TTGAGGCCTTATTCCAGGAATACCAGTGCTATGGAAGTTCGTCAA ACACAAGGGTTCTAAATGAATTAAATTATGATAATGCAGGCACTA ATTTATATAATGAATTAGAAATGAATTATTATGGGAAACAGGAA AATTGGTATAGTCTTAAAAAAAATAGTAGATCACTTGGAGAAAA TGATGATGGAAATAACGAAGACAACGAGAAATTAAGGAAACCAA AACATAAAAAATTAAAGCAACCAGCGGATGGTAATCCTGATCCA AATGCAAACCCAAATGTAGATCCCAATGCCAACCCAAATGTAGA TCCAAATGCAAACCCAAATGTAGATCCAAATGCAAACCCAAATG CAAACCCAAATGCAAACCCAAATGCAAACCCAAATGCAAACCCA AATGCAAACCCAAATGCAAACCCAAATGCAAACCCAAATGCAAA CCCAAATGCAAACCCAAATGCAAACCCAAATGCAAACCCAAATG CAAACCCAAATGCAAACCCAAATGCAAACCCAAATGCAAACCCC AATGCAAATCCTAATGCAAACCCAAATGCAAACCCAAACGTAGA TCCTAATGCAAATCCAAATGCAAACCCAAACGCAAACCCCAATG CAAATCCTAATGCAAACCCCAATGCAAATCCTAATGCAAATCCTA ATGCCAATCCAAATGCAAATCCAAATGCAAACCCAAACGCAAAC CCCAATGCAAATCCTAATGCCAATCCAAATGCAAATCCAAATGCA AACCCAAATGCAAACCCAAATGCAAACCCCAATGCAAATCCTAA TAAAAACAATCAAGGTAATGGACAAGGTCACAATATGCCAAATG ACCCAAACCGAAATGTAGATGAAAATGCTAATGCCAACAGTGCT GTAAAAAATAATAATAACGAAGAACCAAGTGATAAGCACATAAA AGAATATTTAAACAAAATACAAAATTCTCTTTCAACTGAATGGTC CCCATGTAGTGTAACTTGTGGAAATGGTATTCAAGTTAGAATAAA GCCTGGCTCTGCTAATAAACCTAAAGACGAATTAGATTATGCAAA TGATATTGAAAAAAAAATTTGTAAAATGGAAAAATGTTCCAGTGT GTTTAATGTCGTAAATAGTTCAATAGGATTAATAATGGTATTATC CTTCTTGTTCCTTAAT CSP_Soluble ATGTGGTGGCGCCTGTGGTGGCTGCTGCTGCTGCTGCTGCTGCTG  2 TGGCCCATGGTGTGGGCCGAGGCCTTATTCCAGGAATACCAGTGC TATGGAAGTTCGTCAAACACAAGGGTTCTAAATGAATTAAATTAT GATAATGCAGGCACTAATTTATATAATGAATTAGAAATGAATTAT TATGGGAAACAGGAAAATTGGTATAGTCTTAAAAAAAATAGTAG ATCACTTGGAGAAAATGATGATGGAAATAACGAAGACAACGAGA AATTAAGGAAACCAAAACATAAAAAATTAAAGCAACCAGCGGAT GGTAATCCTGATCCAAATGCAAACCCAAATGTAGATCCCAATGCC AACCCAAATGTAGATCCAAATGCAAACCCAAATGTAGATCCAAA TGCAAACCCAAATGCAAACCCAAATGCAAACCCAAATGCAAACC CAAATGCAAACCCAAATGCAAACCCAAATGCAAACCCAAATGCA AACCCAAATGCAAACCCAAATGCAAACCCAAATGCAAACCCAAA TGCAAACCCAAATGCAAACCCAAATGCAAACCCAAATGCAAACC CAAATGCAAACCCCAATGCAAATCCTAATGCAAACCCAAATGCA AACCCAAACGTAGATCCTAATGCAAATCCAAATGCAAACCCAAA CGCAAACCCCAATGCAAATCCTAATGCAAACCCCAATGCAAATC CTAATGCAAATCCTAATGCCAATCCAAATGCAAATCCAAATGCAA ACCCAAACGCAAACCCCAATGCAAATCCTAATGCCAATCCAAAT GCAAATCCAAATGCAAACCCAAATGCAAACCCAAATGCAAACCC CAATGCAAATCCTAATAAAAACAATCAAGGTAATGGACAAGGTC ACAATATGCCAAATGACCCAAACCGAAATGTAGATGAAAATGCT AATGCCAACAGTGCTGTAAAAAATAATAATAACGAAGAACCAAG TGATAAGCACATAAAAGAATATTTAAACAAAATACAAAATTCTCT TTCAACTGAATGGTCCCCATGTAGTGTAACTTGTGGAAATGGTAT TCAAGTTAGAATAAAGCCTGGCTCTGCTAATAAACCTAAAGACG AATTAGATTATGCAAATGATATTGAAAAAAAAATTTGTAAAATG GAAAAATGTTCCAGTGTGTTTAATGTCGTAAATAGTTCAATAGGA TTAATAATGGTATTATCCTTCTTGTTCCTTAAT LSA1_3D7_FL ATGAAACATATTTTGTACATATCATTTTACTTTATCCTTGTTAATT  3 TATTGATATTTCATATAAATGGAAAGATAATAAAGAATTCTGAAA AAGATGAAATCATAAAATCTAACTTGAGAAGTGGTTCTTCAAATT CTAGGAATCGAATAAATGAGGAAAAGCACGAGAAGAAACACGTT TTATCTCATAATTCATATGAGAAAACTAAAAATAATGAAAATAAT AAATTTTTCGATAAGGATAAAGAGTTAACGATGTCTAATGTAAAA AATGTGTCACAAACAAATTTCAAAAGTCTTTTAAGAAATCTTGGT GTTTCAGAGAATATATTCCTTAAAGAAAATAAATTAAATAAGGA AGGGAAATTAATTGAACACATAATAAATGATGATGACGATAAAA AAAAATATATTAAAGGGCAAGACGAAAACAGACAAGAAGATCTT GAACAAGAGAGACTTGCTAAAGAAAAGTTACAGGGGCAACAAA GCGATTTAGAACAAGAGAGACTTGCTAAAGAAAAGTTGCAAGAA CAACAAAGCGATTTAGAACAAGAGAGACTTGCTAAAGAAAAGTT GCAAGAACAACAAAGCGATTTAGAACAAGATAGACTTGCTAAAG AAAAGTTACAAGAGCAACAAAGCGATTTAGAACAAGAGAGACGT GCTAAAGAAAAGTTGCAAGAACAACAAAGCGATTTAGAACGAAC GAAGACTTCTAAAGAAAAGTTACATGAGCAGCAAAGCGATTTAG AACAAGAGAGACGTGCTAAAGAAAAGTTGCAAGAACAACAAAG CGATTTAGAACAAGAGAGACTTGCTAAAGAAAAGTTGCAAGAGC AACAAAGCGATTTAGAACAAGAGAGACTTGCTAAAGAAAAGTTG CAAGAACAACAAAGCGATTTAGAACAAGAGAGACTTGCTAAAGA AAAGTTACAAGAGCAGCAAAGCGATTTAGAACAAGAGAGACTTG CTAAAGAAAAGTTGCAAGAACAACAAAGCGATTTAGAACAAGAG AGACTTGCTAAAGAAAAGTTGCAAGAACAACAAAGCGATTTAGA ACAAGATAGACTTGCTAAAGAAAAGTTACAAGAGCAACAAAGCG ATTTAGAACAAGAGAGACTTGCTAAAGAAAAGTTGCAAGAACAA CAAAGCGATTTAGAACAAGAGAGACGTGCTAAAGAAAAGTTGCA AGAACAACAAAGCGATTTAGAACAAGAGAGACGTGCTAAAGAA AAGTTGCAAGAACAACAAAGCGATTTAGAACAAGAGAGACGTGC TAAAGAAAAGTTGCAAGAACAACAAAGCGATTTAGAACAAGAGA GACGTGCTAAAGAAAAGTTGCAAGAACAACAAAGCGATTTAGAA CAAGAGAGACTTGCTAAAGAAAAGTTGCAAGAACAACAAAGCGA TTTAGAACAAGAGAGACGTGCTAAAGAAAAGTTGCAAGAACAAC AAAGCGATTTAGAACAAGAGAGACTTGCTAAAGAAAAGTTGCAA GAACAACAAAGCGATTTAGAACAAGAGAGACGTGCTAAAGAAA AGTTGCAAGAACAACAAAGCGATTTAGAACAAGAGAGACGTGCT AAAGAAAAGTTGCAAGAACAACAAAGCGATTTAGAACAAGATAG ACTTGCTAAAGAAAAGTTACAAGAGCAACAAAGCGATTTAGAAC AAGAGAGACGTGCTAAAGAAAAGTTGCAAGAACAACAAAGCGA TTTAGAACGAACGAAGGCATCTACAGAAACGTTGCATGAGCAGC AAAGCGATCTTGAACAAGAGAGACTTGCTAAAGAAAAGTTACAA GAGCAGCAAAGCGATTTAGAACAAGAGAGACTTGCTAAAGAAAA GTTACAAGAGCAACAAAGCGATTTAGAACGAACGAAGGCATCTA CAGAAACGTTGCGTGAGCAGCAAAGCGATCTTGAACAAGAGAAA CTAGCTAAAGAAAAGTTACAGGGGCAACAAAGCGATCTTGAACA AGAGAGACTAGCTAAAGAAAAGTTACAGGGGCAACAAAGCGATC TTGAACAAGAGAGACTAGCTAAAGAAAAGTTACAGGGGCAACAA AGCGATCTTGAACAAGAGAGACTAGCTAAAGAAAAGTTACAGGG GCAACAAAGCGATTTAGAACAAGAGAGACTTGCTAAAGAAAAGT TGCAAGAGCGACAAAGCGATTTAGAACAAGAGAGACTTGCTAAA GAAAAGTTGCAAGAACAACAAAGCGATTTAGAACAAGAGAGACT AGCTAAAGAAAAGTTGCAAGAACAACAAAGCGATTTAGAACAAG ACAGACTTGCTAAAGAAAAGTTGCAAGAACAACAAAGCGATTTA GAACAAGAGAGACTAGCTAAGGAAAAGTTACAGGGGCAGCATA GCGATTTAGAACGAACGAAGGCATCTAAAGAAACGTTGCAAGAA CAACAAAGCGATTTAGAACAAGAGAGACTTGCTAAAGAAAAGTT GCAAGAACAACAAAGCGATTTAGAACAAGAGAGACGTGCTAAAG AAAAGTTGCAAGAACAACAAAGCGATTTAGAACAAGAGAGACGT GCTAAAGAAAAGTTGCAAGAACAACAAAGCGATTTAGAACAAGA GAGACGTGCTAAAGAAAAGTTGCAAGAGCAGCAAAGAGATTTAG AACAAAGGAAGGCTGATACGAAAAAAAATTTAGAAAGAAAAAA GGAACATGGAGATGTATTAGCAGAGGATTTATATGGTCGTTTAGA AATACCAGCTATAGAACTTCCATCAGAAAATGAACGTGGATATTA TATACCACATCAATCTTCTTTACCTCAGGACAACAGAGGGAATAG TAGAGATTCGAAGGAAATATCTATAATAGAAAATACAAATAGAG AATCTATTACAACAAATGTTGAAGGACGAAGGGATATACATAAA GGACATCTTGAAGAAAAGAAAGATGGTTCAATAAAACCAGAACA AAAAGAAGATAAATCTGCTGACATACAAAATCATACATTAGAGA CAGTAAATATTTCTGATGTTAATGATTTTCAAATAAGTAAGTATG AGGATGAAATAAGTGCTGAATATGACGATTCATTAATAGATGAA GAAGAAGATGATGAAGACTTAGACGAATTTAAGCCTATTGTGCA ATATGACAATTTCCAAGATGAAGAAAACATAGGAATTTATAAAG AACTAGAAGATTTGATAGAGAAAAATGAAAATTTAGATGATTTA GATGAAGGAATAGAAAAATCATCAGAAGAATTATCTGAAGAAAA AATAAAAAAAGGAAAGAAATATGAAAAAACAAAGGATAATAAT TTTAAACCAAATGATAAAAGTTTGTATGATGAGCATATTAAAAAA TATAAAAATGATAAGCAGGTTAATAAGGAAAAGGAAAAATTCAT AAAATCATTGTTTCATATATTTGACGGAGACAATGAAATTTTACA GATCGTGGATGAGTTATCTGAAGATATAACTAAATATTTTATGAA ACTA MSP1_3D7_FL ATGAAGATCATATTCTTTTTATGTTCATTTCTTTTTTTTATTATAAA  4 TACACAATGTGTAACACATGAAAGTTATCAAGAACTTGTCAAAA AACTAGAAGCTTTAGAAGATGCAGTATTGACAGGTTATAGTTTAT TTCAAAAGGAAAAAATGGTATTAAATGAAGAAGAAATTACTACA AAAGGTGCAAGTGCTCAAAGTGGTGCAAGTGCTCAAAGTGGTGC AAGTGCTCAAAGTGGTGCAAGTGCTCAAAGTGGTGCAAGTGCTC AAAGTGGTGCAAGTGCTCAAAGTGGTACAAGTGGTCCAAGTGGT CCAAGTGGTACAAGTCCATCATCTCGTTCAAACACTTTACCTCGT TCAAATACTTCATCTGGTGCAAGCCCTCCAGCTGATGCAAGCGAT TCAGATGCTAAATCTTACGCTGATTTAAAACACAGAGTACGAAAT TACTTGTTCACTATTAAAGAACTCAAATATCCCGAACTCTTTGATT TAACCAATCATATGTTAACTTTGTGTGATAATATTCATGGTTTCAA ATATTTAATTGATGGATATGAAGAAATTAATGAATTATTATATAA ATTAAACTTTTATTTTGATTTATTAAGAGCAAAATTAAATGATGT ATGTGCTAATGATTATTGTCAAATACCTTTCAATCTTAAAATTCGT GCAAATGAATTAGACGTACTTAAAAAACTTGTGTTCGGATATAGA AAACCATTAGACAATATTAAAGATAATGTAGGAAAAATGGAAGA TTACATTAAAAAAAATAAAACAACCATAGCAAATATAAATGAAT TAATTGAAGGAAGTAAGAAAACAATTGATCAAAATAAGAATGCA GATAATGAAGAAGGGAAAAAAAAATTATACCAAGCTCAATATGA TCTTTCTATTTACAATAAACAATTAGAAGAAGCACATAATTTAAT AAGCGTTTTAGAAAAACGTATTGACACTTTAAAAAAAAATGAAA ACATAAAGAAATTACTTGATAAGATAAATGAAATTAAAAATCCC CCACCGGCCAATTCTGGAAATACACCAAATACTCTCCTTGATAAG AACAAAAAAATCGAGGAACACGAAGAAAAAATAAAAGAAATTG CCAAAACTATTAAATTTAACATTGATAGTTTATTTACTGATCCACT TGAATTAGAATATTATTTAAGAGAAAAAAATAAAAAAGTTGATG TAACACCTAAATCACAAGATCCTACGAAATCTGTTCAAATACCAA AAGTTCCTTATCCAAATGGTATTGTATATCCTTTACCACTCACTGA TATTCATAATTCATTAGCTGCAGATAATGATAAAAATTCATATGG TGATTTAATGAATCCTCATACTAAAGAAAAAATTAATGAAAAAAT TATTACAGATAATAAGGAAAGAAAAATATTCATTAATAACATTA AAAAAAAAATTGATTTAGAAGAAAAAAACATTAATCACACAAAA GAACAAAATAAAAAATTACTTGAAGATTATGAAAAGTCAAAAAA GGATTATGAAGAATTACTTGAAAAATTTTATGAAATGAAATTTAA TAATAATTTTAACAAAGATGTCGTAGATAAAATATTCAGTGCAAG ATATACATATAATGTTGAAAAACAAAGATATAATAATAAATTTTC ATCCTCTAATAATTCTGTATATAATGTTCAAAAATTAAAAAAGGC TCTTTCATATCTTGAAGATTATTCTTTAAGAAAAGGAATTTCTGAA AAAGATTTTAATCATTATTATACTTTGAAAACTGGCCTCGAAGCT GATATAAAAAAATTAACAGAAGAAATAAAGAGTAGTGAAAACA AAATTCTTGAAAAAAATTTTAAAGGACTAACACATTCAGCAAATG GTTCCTTAGAAGTATCTGATATTGTAAAATTACAAGTACAAAAAG TTTTATTAATTAAAAAAATAGAAGACTTAAGAAAGATAGAATTAT TTTTAAAAAATGCACAACTAAAAGATAGTATTCATGTACCAAATA TTTATAAACCACAAAATAAACCAGAACCATATTATTTAATTGTAT TAAAAAAAGAAGTAGATAAATTAAAAGAATTTATACCAAAAGTA AAAGACATGTTAAAGAAAGAACAAGCTGTCTTATCAAGTATTAC ACAACCTTTAGTTGCAGCAAGCGAAACAACTGAAGATGGGGGTC ACTCCACACACACATTATCCCAATCAGGAGAAACAGAAGTAACA GAAGAAACAGAAGAAACAGAAGAAACAGTAGGACACACAACAA CGGTAACAATAACATTACCACCAACACAACCATCACCACCAAAA GAAGTAAAAGTTGTTGAAAATTCAATAGAACATAAGAGTAATGA CAATTCACAAGCCTTGACAAAAACAGTTTATCTAAAGAAATTAGA TGAATTTTTAACTAAATCATATATATGTCATAAATATATTTTAGTA TCAAACTCTAGTATGGACCAAAAATTATTAGAGGTATATAATCTT ACTCCAGAAGAAGAAAATGAATTAAAATCATGTGATCCATTAGA TTTATTATTTAATATTCAAAATAACATACCTGCTATGTATTCATTA TATGATAGTATGAACAATGATTTACAACATCTCTTTTTTGAATTAT ATCAAAAGGAAATGATTTATTATTTACATAAACTAAAAGAGGAA AATCACATCAAAAAATTATTAGAGGAGCAAAAACAAATAACTGG AACATCATCTACATCCAGTCCTGGAAATACAACCGTAAATACTGC TCAATCCGCAACTCACAGTAATTCCCAAAACCAACAATCAAATGC ATCCTCTACCAATACCCAAAATGGTGTAGCTGTATCATCTGGTCC TGCTGTAGTTGAAGAAAGTCATGATCCCTTAACAGTATTGTCTAT TAGTAACGATTTGAAAGGTATTGTTAGTCTCTTAAATCTTGGAAA TAAAACTAAAGTACCTAATCCATTAACCATTTCTACAACAGAGAT GGAAAAATTTTATGAGAATATTTTAAAAAATAATGATACCTATTT TAATGATGATATCAAACAATTCGTAAAATCTAATTCAAAAGTAAT TACAGGTTTGACCGAAACACAAAAAAATGCATTAAATGATGAAA TTAAAAAATTAAAAGATACTTTACAGTTATCATTTGATTTATATA ATAAATATAAATTAAAATTAGATAGATTATTTAATAAGAAAAAA GAACTTGGCCAAGACAAAATGCAAATTAAAAAACTTACTTTATTA AAAGAACAATTAGAATCAAAATTGAATTCACTTAATAACCCACAT AATGTATTACAAAACTTTTCTGTTTTCTTTAACAAAAAAAAAGAA GCTGAAATAGCAGAAACTGAAAACACATTAGAAAACACAAAAAT ATTATTGAAACATTATAAAGGACTTGTTAAATATTATAATGGTGA ATCATCTCCATTAAAAACTTTAAGTGAAGTATCAATTCAAACAGA AGATAATTATGCCAATTTAGAAAAATTTAGAGTATTAAGTAAAAT AGATGGAAAACTCAATGATAATTTACATTTAGGAAAGAAAAAAT TATCTTTCTTATCAAGTGGATTACATCATTTAATTACTGAATTAAA AGAAGTAATAAAAAATAAAAATTATACAGGTAATTCTCCAAGTG AAAATAATAAGAAAGTTAACGAAGCTTTAAAATCTTACGAAAAT TTTCTCCCAGAAGCAAAAGTTACAACAGTTGTAACTCCACCTCAA CCAGATGTAACTCCATCTCCATTATCTGTAAGGGTAAGTGGTAGT TCAGGATCCACAAAAGAAGAAACACAAATACCAACTTCAGGCTC TTTATTAACAGAATTACAACAAGTAGTACAATTACAAAATTATGA CGAAGAAGATGATTCCTTAGTTGTATTACCCATTTTTGGAGAATC CGAAGATAATGACGAATATTTAGATCAAGTAGTAACTGGAGAAG CAATATCTGTCACAATGGATAATATCCTCTCAGGATTTGAAAATG AATATGATGTTATATATTTAAAACCTTTAGCTGGAGTATATAGAA GCTTAAAAAAACAAATTGAAAAAAACATTTTTACATTTAATTTAA ATTTGAACGATATCTTAAATTCACGTCTTAAGAAACGAAAATATT TCTTAGATGTATTAGAATCTGATTTAATGCAATTTAAACATATATC CTCAAATGAATACATTATTGAAGATTCATTTAAATTATTGAATTC AGAACAAAAAAACACACTTTTAAAAAGTTACAAATATATAAAAG AATCAGTAGAAAATGATATTAAATTTGCACAGGAAGGTATAAGT TATTATGAAAAGGTTTTAGCGAAATATAAGGATGATTTAGAATCA ATTAAAAAAGTTATCAAAGAAGAAAAGGAGAAGTTCCCATCATC ACCACCAACAACACCTCCGTCACCAGCAAAAACAGACGAACAAA AGAAGGAAAGTAAGTTCCTTCCATTTTTAACAAACATTGAGACCT TATACAATAACTTAGTTAATAAAATTGACGATTACTTAATTAACT TAAAGGCAAAGATTAACGATTGTAATGTTGAAAAAGATGAAGCA CATGTTAAAATAACTAAACTTAGTGATTTAAAAGCAATTGATGAC AAAATAGATCTTTTTAAAAACCCTTACGACTTCGAAGCAATTAAA AAATTGATAAATGATGATACGAAAAAAGATATGCTTGGCAAATT ACTTAGTACAGGATTAGTTCAAAATTTTCCTAATACAATAATATC AAAATTAATTGAAGGAAAATTCCAAGATATGTTAAACATTTCACA ACACCAATGCGTAAAAAAACAATGTCCAGAAAATTCTGGATGTTT CAGACATTTAGATGAAAGAGAAGAATGTAAATGTTTATTAAATTA CAAACAAGAAGGTGATAAATGTGTTGAAAATCCAAATCCTACTT GTAACGAAAATAATGGTGGATGTGATGCAGATGCCACATGTACC GAAGAAGATTCAGGTAGCAGCAGAAAGAAAATCACATGTGAATG TACTAAACCTGATTCTTATCCACTTTTCGATGGTATTTTCTGCAGT TCCTCTAACTTCTTAGGAATATCATTCTTATTAATACTCATGTTAA TATTATACAGTTTCATT AMA1_3D7_FL_ ATGAGAAAATTATACTGCGTATTATTATTGAGCGCCTTTGAGTTT  5 DX ACATATATGATAAACTTTGGAAGAGGACAGAATTATTGGGAACA TCCATATCAAAATAGTGATGTGTATCGTCCAATCAACGAACATAG GGAACATCCAAAAGAATACGAATATCCATTACACCAGGAACATA CATACCAACAAGAAGATTCAGGAGAAGACGAAAATACATTACAA CACGCATATCCAATAGACCACGAAGGTGCTGAACCCGCACCACA AGAACAAAACTTATTCTCAAGCATTGAAATAGTCGAACGGTCGA ATTATATGGGTAATCCGTGGACGGAATACATGGCAAAATATGAT ATTGAAGAAGTTCATGGTTCAGGTATAAGGGTAGATTTAGGAGA GGATGCTGAAGTAGCTGGAACTCAATACAGACTTCCATCAGGTA AATGTCCAGTATTCGGTAAAGGTATAATTATTGAGAATTCAAATA CTACTTTTCTAACACCGGTAGCTACAGGAAATCAATATCTAAAAG ATGGAGGGTTTGCGTTTCCTCCAACAGAACCTCTTATGTCACCAA TGACACTAGATGAAATGAGACATTTTTATAAGGATAATAAATACG TAAAAAACCTCGACGAATTGACTTTATGTTCAAGACATGCAGGGA ATATGATTCCAGATAATGATAAAAACTCAAATTATAAATACCCGG CTGTTTATGATGATAAAGATAAAAAGTGTCATATCCTGTATATTG CAGCTCAAGAAAATAATGGTCCTAGATATTGTAATAAAGACGAA AGTAAAAGAAACAGCATGTTTTGTTTCAGACCAGCAAAAGACAT AAGCTTCCAAAACTATACTTACTTGTCTAAAAATGTAGTTGATAA CTGGGAAAAAGTCTGCCCTAGAAAGAATTTACAGAATGCGAAAT TCGGACTATGGGTCGATGGTAACTGTGAAGATATACCACATGTAA ATGAATTTCCAGCAATAGATCTTTTCGAATGCAATAAACTGGTTT TTGAATTGAGTGCCTCGGATCAGCCAAAACAGTATGAACAACATT TAACAGATTACGAAAAAATCAAAGAAGGTTTTAAAAATAAAAAT GCTAGTATGATCAAGAGTGCTTTTCTTCCCACGGGTGCTTTTAAG GCAGATCGTTATAAAAGTCACGGTAAAGGATATAATTGGGGTAA CTATAACACAGAGACACAAAAGTGTGAAATTTTTAATGTTAAACC AACATGTCTCATAAACAATTCCTCATACATTGCTACCACTGCTTTA TCCCATCCCATCGAAGTAGAGAACAATTTTCCTTGTAGCCTATAC AAAGATGAAATAATGAAAGAAATCGAGAGAGAATCAAAACGAA TTAAATTAAATGATAACGATGACGAGGGGAACAAAAAAATTATA GCTCCACGCATTTTTATTTCAGACGATAAAGACTCGTTAAAATGC CCATGTGACCCTGAAATGGTCAGTAACAGTACATGCCGCTTCTTT GTATGTAAATGTGTAGAAAGGAGGGCAGAAGTAACATCAAATAA CGAGGTTGTAGTAAAAGAAGAATACAAAGATGAGTACGCAGATA TTCCCGAACACAAACCAACCTATGATAAGATGAAAATAATAATA GCATCATCAGCTGCAGTTGCTGTACTAGCGACGATTTTAATGGTT TATCTTTATAAAAGAAAGGGAAATGCTGAAAAATATGATAAAAT GGATGAACCGCAAGATTATGGGAAATCCAATTCACGAAACGATG AGATGCTTGATCCTGAGGCATCTTTCTGGGGGGAAGAAAAAAGA GCATCACATACAACACCAGTACTGATGGAAAAACCATACTAT PF3D7_1335900 | ATGAATCATCTTGGGAATGTTAAATATTTAGTCATTGTGTTTTTGA  6 Plasmodium TTTTCTTTGATTTGTTTCTAGTTAATGGTAGAGATGTGCAAAACAA falciparum 3D7 | TATAGTGGATGAAATAAAATATCGTGAAGAAGTATGTAATGATG thrombospondin- AGGTAGATCTTTACCTTCTAATGGATTGTTCTGGAAGTATACGTC related GTCATAATTGGGTGAACCATGCAGTACCTCTAGCTATGAAATTGA anonymous TACAACAATTAAATCTTAATGATAATGCAATTCACTTATATGCTA protein (TRAP) | GTGTTTTTTCAAACAATGCAAGAGAAATTATTAGATTACATAGTG CDS | ATGCATCTAAAAACAAAGAGAAGGCTTTAATTATTATAAAGTCAC length = 1725 TCTTAAGTACAAATCTTCCATATGGTAAAACAAACTTAACTGATG CACTGTTACAAGTAAGAAAACATTTAAATGACCGAATCAATAGA GAGAATGCTAATCAATTAGTTGTTATATTAACAGATGGAATTCCA GATAGTATTCAAGATTCATTAAAAGAATCAAGAAAATTAAGTGA TCGTGGTGTTAAAATAGCTGTTTTTGGTATTGGACAAGGTATTAA TGTAGCTTTCAACAGATTTCTTGTAGGTTGTCATCCATCAGATGGT AAATGTAACTTGTATGCTGATTCTGCATGGGAAAATGTAAAAAAT GTTATCGGACCCTTTATGAAGGCTGTTTGTGTTGAAGTAGAAAAA ACAGCAAGTTGTGGTGTTTGGGACGAATGGTCTCCATGTAGTGTA ACTTGTGGTAAAGGTACCAGGTCAAGAAAAAGAGAAATCTTACA CGAAGGATGTACAAGTGAATTACAAGAACAATGTGAAGAAGAAA GATGTCTTCCAAAACGGGAACCATTAGATGTTCCAGATGAACCCG AAGATGATCAACCTAGACCAAGAGGAGATAATTTTGCTGTCGAA AAACCAAACGAAAATATAATAGATAATAATCCACAAGAACCTTC ACCAAATCCAGAAGAAGGAAAGGGTGAAAATCCAAACGGATTTG ATTTAGATGAAAATCCAGAAAATCCACCAAATCCACCAAATCCA CCAAATCCACCAAATCCACCAAATCCACCAAATCCAGATATTCCT GAACAAGAACCAAATATACCTGAAGATTCAGAAAAAGAAGTACC TTCTGATGTTCCAAAAAATCCAGAAGACGATCGAGAAGAAAACT TTGATATTCCAAAGAAACCCGAAAATAAGCACGATAATCAAAAT AATTTACCAAATGATAAAAGTGATAGATATATTCCATATTCACCA TTATCTCCAAAAGTTTTGGATAATGAAAGGAAACAAAGTGACCCC CAAAGTCAAGATAATAATGGAAATAGGCACGTACCTAATAGTGA AGATAGAGAAACACGTCCACATGGTAGAAATAATGAAAATAGAT CATACAATAGAAAACATAACAATACTCCAAAACATCCTGAAAGG GAAGAACATGAAAAGCCAGATAATAATAAAAAAAAAGCAGGAT CAGATAATAAATATAAAATTGCAGGTGGAATAGCTGGAGGATTA GCTTTACTCGCATGTGCTGGACTTGCTTATAAATTCGTAGTACCA GGAGCAGCAACACCCTATGCCGGAGAACCTGCACCTTTTGATGA AACATTAGGTGAAGAAGATAAAGATTTGGACGAACCTGAACAAT TCAGATTACCTGAAGAAAACGAGTGGAATTAA Plasmodium mRNA Sequences CSP_Full_length AUGAUGAGAAAAUUAGCUAUUUUAUCUGUUUCUUCCUUUUUAU  7 UUGUUGAGGCCUUAUUCCAGGAAUACCAGUGCUAUGGAAGUUC GUCAAACACAAGGGUUCUAAAUGAAUUAAAUUAUGAUAAUGCA GGCACUAAUUUAUAUAAUGAAUUAGAAAUGAAUUAUUAUGGG AAACAGGAAAAUUGGUAUAGUCUUAAAAAAAAUAGUAGAUCAC UUGGAGAAAAUGAUGAUGGAAAUAACGAAGACAACGAGAAAU UAAGGAAACCAAAACAUAAAAAAUUAAAGCAACCAGCGGAUGG UAAUCCUGAUCCAAAUGCAAACCCAAAUGUAGAUCCCAAUGCC AACCCAAAUGUAGAUCCAAAUGCAAACCCAAAUGUAGAUCCAA AUGCAAACCCAAAUGCAAACCCAAAUGCAAACCCAAAUGCAAA CCCAAAUGCAAACCCAAAUGCAAACCCAAAUGCAAACCCAAAU GCAAACCCAAAUGCAAACCCAAAUGCAAACCCAAAUGCAAACC CAAAUGCAAACCCAAAUGCAAACCCAAAUGCAAACCCAAAUGC AAACCCAAAUGCAAACCCCAAUGCAAAUCCUAAUGCAAACCCA AAUGCAAACCCAAACGUAGAUCCUAAUGCAAAUCCAAAUGCAA ACCCAAACGCAAACCCCAAUGCAAAUCCUAAUGCAAACCCCAAU GCAAAUCCUAAUGCAAAUCCUAAUGCCAAUCCAAAUGCAAAUC CAAAUGCAAACCCAAACGCAAACCCCAAUGCAAAUCCUAAUGC CAAUCCAAAUGCAAAUCCAAAUGCAAACCCAAAUGCAAACCCA AAUGCAAACCCCAAUGCAAAUCCUAAUAAAAACAAUCAAGGUA AUGGACAAGGUCACAAUAUGCCAAAUGACCCAAACCGAAAUGU AGAUGAAAAUGCUAAUGCCAACAGUGCUGUAAAAAAUAAUAAU AACGAAGAACCAAGUGAUAAGCACAUAAAAGAAUAUUUAAACA AAAUACAAAAUUCUCUUUCAACUGAAUGGUCCCCAUGUAGUGU AACUUGUGGAAAUGGUAUUCAAGUUAGAAUAAAGCCUGGCUCU GCUAAUAAACCUAAAGACGAAUUAGAUUAUGCAAAUGAUAUUG AAAAAAAAAUUUGUAAAAUGGAAAAAUGUUCCAGUGUGUUUA AUGUCGUAAAUAGUUCAAUAGGAUUAAUAAUGGUAUUAUCCUU CUUGUUCCUUAAU CSP_Soluble AUGUGGUGGCGCCUGUGGUGGCUGCUGCUGCUGCUGCUGCUGC  8 UGUGGCCCAUGGUGUGGGCCGAGGCCUUAUUCCAGGAAUACCA GUGCUAUGGAAGUUCGUCAAACACAAGGGUUCUAAAUGAAUUA AAUUAUGAUAAUGCAGGCACUAAUUUAUAUAAUGAAUUAGAA AUGAAUUAUUAUGGGAAACAGGAAAAUUGGUAUAGUCUUAAA AAAAAUAGUAGAUCACUUGGAGAAAAUGAUGAUGGAAAUAAC GAAGACAACGAGAAAUUAAGGAAACCAAAACAUAAAAAAUUAA AGCAACCAGCGGAUGGUAAUCCUGAUCCAAAUGCAAACCCAAA UGUAGAUCCCAAUGCCAACCCAAAUGUAGAUCCAAAUGCAAAC CCAAAUGUAGAUCCAAAUGCAAACCCAAAUGCAAACCCAAAUG CAAACCCAAAUGCAAACCCAAAUGCAAACCCAAAUGCAAACCC AAAUGCAAACCCAAAUGCAAACCCAAAUGCAAACCCAAAUGCA AACCCAAAUGCAAACCCAAAUGCAAACCCAAAUGCAAACCCAA AUGCAAACCCAAAUGCAAACCCAAAUGCAAACCCCAAUGCAAA UCCUAAUGCAAACCCAAAUGCAAACCCAAACGUAGAUCCUAAU GCAAAUCCAAAUGCAAACCCAAACGCAAACCCCAAUGCAAAUC CUAAUGCAAACCCCAAUGCAAAUCCUAAUGCAAAUCCUAAUGC CAAUCCAAAUGCAAAUCCAAAUGCAAACCCAAACGCAAACCCC AAUGCAAAUCCUAAUGCCAAUCCAAAUGCAAAUCCAAAUGCAA ACCCAAAUGCAAACCCAAAUGCAAACCCCAAUGCAAAUCCUAA UAAAAACAAUCAAGGUAAUGGACAAGGUCACAAUAUGCCAAAU GACCCAAACCGAAAUGUAGAUGAAAAUGCUAAUGCCAACAGUG CUGUAAAAAAUAAUAAUAACGAAGAACCAAGUGAUAAGCACAU AAAAGAAUAUUUAAACAAAAUACAAAAUUCUCUUUCAACUGAA UGGUCCCCAUGUAGUGUAACUUGUGGAAAUGGUAUUCAAGUUA GAAUAAAGCCUGGCUCUGCUAAUAAACCUAAAGACGAAUUAGA UUAUGCAAAUGAUAUUGAAAAAAAAAUUUGUAAAAUGGAAAA AUGUUCCAGUGUGUUUAAUGUCGUAAAUAGUUCAAUAGGAUUA AUAAUGGUAUUAUCCUUCUUGUUCCUUAAU LSA1_3D7_FL AUGAAACAUAUUUUGUACAUAUCAUUUUACUUUAUCCUUGUUA  9 AUUUAUUGAUAUUUCAUAUAAAUGGAAAGAUAAUAAAGAAUU CUGAAAAAGAUGAAAUCAUAAAAUCUAACUUGAGAAGUGGUUC UUCAAAUUCUAGGAAUCGAAUAAAUGAGGAAAAGCACGAGAAG AAACACGUUUUAUCUCAUAAUUCAUAUGAGAAAACUAAAAAUA AUGAAAAUAAUAAAUUUUUCGAUAAGGAUAAAGAGUUAACGA UGUCUAAUGUAAAAAAUGUGUCACAAACAAAUUUCAAAAGUCU UUUAAGAAAUCUUGGUGUUUCAGAGAAUAUAUUCCUUAAAGAA AAUAAAUUAAAUAAGGAAGGGAAAUUAAUUGAACACAUAAUA AAUGAUGAUGACGAUAAAAAAAAAUAUAUUAAAGGGCAAGAC GAAAACAGACAAGAAGAUCUUGAACAAGAGAGACUUGCUAAAG AAAAGUUACAGGGGCAACAAAGCGAUUUAGAACAAGAGAGACU UGCUAAAGAAAAGUUGCAAGAACAACAAAGCGAUUUAGAACAA GAGAGACUUGCUAAAGAAAAGUUGCAAGAACAACAAAGCGAUU UAGAACAAGAUAGACUUGCUAAAGAAAAGUUACAAGAGCAACA AAGCGAUUUAGAACAAGAGAGACGUGCUAAAGAAAAGUUGCAA GAACAACAAAGCGAUUUAGAACGAACGAAGACUUCUAAAGAAA AGUUACAUGAGCAGCAAAGCGAUUUAGAACAAGAGAGACGUGC UAAAGAAAAGUUGCAAGAACAACAAAGCGAUUUAGAACAAGAG AGACUUGCUAAAGAAAAGUUGCAAGAGCAACAAAGCGAUUUAG AACAAGAGAGACUUGCUAAAGAAAAGUUGCAAGAACAACAAAG CGAUUUAGAACAAGAGAGACUUGCUAAAGAAAAGUUACAAGAG CAGCAAAGCGAUUUAGAACAAGAGAGACUUGCUAAAGAAAAGU UGCAAGAACAACAAAGCGAUUUAGAACAAGAGAGACUUGCUAA AGAAAAGUUGCAAGAACAACAAAGCGAUUUAGAACAAGAUAGA CUUGCUAAAGAAAAGUUACAAGAGCAACAAAGCGAUUUAGAAC AAGAGAGACUUGCUAAAGAAAAGUUGCAAGAACAACAAAGCGA UUUAGAACAAGAGAGACGUGCUAAAGAAAAGUUGCAAGAACAA CAAAGCGAUUUAGAACAAGAGAGACGUGCUAAAGAAAAGUUGC AAGAACAACAAAGCGAUUUAGAACAAGAGAGACGUGCUAAAGA AAAGUUGCAAGAACAACAAAGCGAUUUAGAACAAGAGAGACGU GCUAAAGAAAAGUUGCAAGAACAACAAAGCGAUUUAGAACAAG AGAGACUUGCUAAAGAAAAGUUGCAAGAACAACAAAGCGAUUU AGAACAAGAGAGACGUGCUAAAGAAAAGUUGCAAGAACAACAA AGCGAUUUAGAACAAGAGAGACUUGCUAAAGAAAAGUUGCAAG AACAACAAAGCGAUUUAGAACAAGAGAGACGUGCUAAAGAAAA GUUGCAAGAACAACAAAGCGAUUUAGAACAAGAGAGACGUGCU AAAGAAAAGUUGCAAGAACAACAAAGCGAUUUAGAACAAGAUA GACUUGCUAAAGAAAAGUUACAAGAGCAACAAAGCGAUUUAGA ACAAGAGAGACGUGCUAAAGAAAAGUUGCAAGAACAACAAAGC GAUUUAGAACGAACGAAGGCAUCUACAGAAACGUUGCAUGAGC AGCAAAGCGAUCUUGAACAAGAGAGACUUGCUAAAGAAAAGUU ACAAGAGCAGCAAAGCGAUUUAGAACAAGAGAGACUUGCUAAA GAAAAGUUACAAGAGCAACAAAGCGAUUUAGAACGAACGAAGG CAUCUACAGAAACGUUGCGUGAGCAGCAAAGCGAUCUUGAACA AGAGAAACUAGCUAAAGAAAAGUUACAGGGGCAACAAAGCGAU CUUGAACAAGAGAGACUAGCUAAAGAAAAGUUACAGGGGCAAC AAAGCGAUCUUGAACAAGAGAGACUAGCUAAAGAAAAGUUACA GGGGCAACAAAGCGAUCUUGAACAAGAGAGACUAGCUAAAGAA AAGUUACAGGGGCAACAAAGCGAUUUAGAACAAGAGAGACUUG CUAAAGAAAAGUUGCAAGAGCGACAAAGCGAUUUAGAACAAGA GAGACUUGCUAAAGAAAAGUUGCAAGAACAACAAAGCGAUUUA GAACAAGAGAGACUAGCUAAAGAAAAGUUGCAAGAACAACAAA GCGAUUUAGAACAAGACAGACUUGCUAAAGAAAAGUUGCAAGA ACAACAAAGCGAUUUAGAACAAGAGAGACUAGCUAAGGAAAAG UUACAGGGGCAGCAUAGCGAUUUAGAACGAACGAAGGCAUCUA AAGAAACGUUGCAAGAACAACAAAGCGAUUUAGAACAAGAGAG ACUUGCUAAAGAAAAGUUGCAAGAACAACAAAGCGAUUUAGAA CAAGAGAGACGUGCUAAAGAAAAGUUGCAAGAACAACAAAGCG AUUUAGAACAAGAGAGACGUGCUAAAGAAAAGUUGCAAGAACA ACAAAGCGAUUUAGAACAAGAGAGACGUGCUAAAGAAAAGUUG CAAGAGCAGCAAAGAGAUUUAGAACAAAGGAAGGCUGAUACGA AAAAAAAUUUAGAAAGAAAAAAGGAACAUGGAGAUGUAUUAG CAGAGGAUUUAUAUGGUCGUUUAGAAAUACCAGCUAUAGAACU UCCAUCAGAAAAUGAACGUGGAUAUUAUAUACCACAUCAAUCU UCUUUACCUCAGGACAACAGAGGGAAUAGUAGAGAUUCGAAGG AAAUAUCUAUAAUAGAAAAUACAAAUAGAGAAUCUAUUACAAC AAAUGUUGAAGGACGAAGGGAUAUACAUAAAGGACAUCUUGAA GAAAAGAAAGAUGGUUCAAUAAAACCAGAACAAAAAGAAGAUA AAUCUGCUGACAUACAAAAUCAUACAUUAGAGACAGUAAAUAU UUCUGAUGUUAAUGAUUUUCAAAUAAGUAAGUAUGAGGAUGA AAUAAGUGCUGAAUAUGACGAUUCAUUAAUAGAUGAAGAAGA AGAUGAUGAAGACUUAGACGAAUUUAAGCCUAUUGUGCAAUAU GACAAUUUCCAAGAUGAAGAAAACAUAGGAAUUUAUAAAGAAC UAGAAGAUUUGAUAGAGAAAAAUGAAAAUUUAGAUGAUUUAG AUGAAGGAAUAGAAAAAUCAUCAGAAGAAUUAUCUGAAGAAA AAAUAAAAAAAGGAAAGAAAUAUGAAAAAACAAAGGAUAAUA AUUUUAAACCAAAUGAUAAAAGUUUGUAUGAUGAGCAUAUUA AAAAAUAUAAAAAUGAUAAGCAGGUUAAUAAGGAAAAGGAAA AAUUCAUAAAAUCAUUGUUUCAUAUAUUUGACGGAGACAAUGA AAUUUUACAGAUCGUGGAUGAGUUAUCUGAAGAUAUAACUAAA UAUUUUAUGAAACUA MSP1_3D7_FL AUGAAGAUCAUAUUCUUUUUAUGUUCAUUUCUUUUUUUUAUUA 10 UAAAUACACAAUGUGUAACACAUGAAAGUUAUCAAGAACUUGU CAAAAAACUAGAAGCUUUAGAAGAUGCAGUAUUGACAGGUUAU AGUUUAUUUCAAAAGGAAAAAAUGGUAUUAAAUGAAGAAGAA AUUACUACAAAAGGUGCAAGUGCUCAAAGUGGUGCAAGUGCUC AAAGUGGUGCAAGUGCUCAAAGUGGUGCAAGUGCUCAAAGUGG UGCAAGUGCUCAAAGUGGUGCAAGUGCUCAAAGUGGUACAAGU GGUCCAAGUGGUCCAAGUGGUACAAGUCCAUCAUCUCGUUCAA ACACUUUACCUCGUUCAAAUACUUCAUCUGGUGCAAGCCCUCC AGCUGAUGCAAGCGAUUCAGAUGCUAAAUCUUACGCUGAUUUA AAACACAGAGUACGAAAUUACUUGUUCACUAUUAAAGAACUCA AAUAUCCCGAACUCUUUGAUUUAACCAAUCAUAUGUUAACUUU GUGUGAUAAUAUUCAUGGUUUCAAAUAUUUAAUUGAUGGAUA UGAAGAAAUUAAUGAAUUAUUAUAUAAAUUAAACUUUUAUUU UGAUUUAUUAAGAGCAAAAUUAAAUGAUGUAUGUGCUAAUGA UUAUUGUCAAAUACCUUUCAAUCUUAAAAUUCGUGCAAAUGAA UUAGACGUACUUAAAAAACUUGUGUUCGGAUAUAGAAAACCAU UAGACAAUAUUAAAGAUAAUGUAGGAAAAAUGGAAGAUUACA UUAAAAAAAAUAAAACAACCAUAGCAAAUAUAAAUGAAUUAAU UGAAGGAAGUAAGAAAACAAUUGAUCAAAAUAAGAAUGCAGA UAAUGAAGAAGGGAAAAAAAAAUUAUACCAAGCUCAAUAUGAU CUUUCUAUUUACAAUAAACAAUUAGAAGAAGCACAUAAUUUAA UAAGCGUUUUAGAAAAACGUAUUGACACUUUAAAAAAAAAUGA AAACAUAAAGAAAUUACUUGAUAAGAUAAAUGAAAUUAAAAA UCCCCCACCGGCCAAUUCUGGAAAUACACCAAAUACUCUCCUUG AUAAGAACAAAAAAAUCGAGGAACACGAAGAAAAAAUAAAAGA AAUUGCCAAAACUAUUAAAUUUAACAUUGAUAGUUUAUUUACU GAUCCACUUGAAUUAGAAUAUUAUUUAAGAGAAAAAAAUAAA AAAGUUGAUGUAACACCUAAAUCACAAGAUCCUACGAAAUCUG UUCAAAUACCAAAAGUUCCUUAUCCAAAUGGUAUUGUAUAUCC UUUACCACUCACUGAUAUUCAUAAUUCAUUAGCUGCAGAUAAU GAUAAAAAUUCAUAUGGUGAUUUAAUGAAUCCUCAUACUAAAG AAAAAAUUAAUGAAAAAAUUAUUACAGAUAAUAAGGAAAGAA AAAUAUUCAUUAAUAACAUUAAAAAAAAAAUUGAUUUAGAAG AAAAAAACAUUAAUCACACAAAAGAACAAAAUAAAAAAUUACU UGAAGAUUAUGAAAAGUCAAAAAAGGAUUAUGAAGAAUUACU UGAAAAAUUUUAUGAAAUGAAAUUUAAUAAUAAUUUUAACAA AGAUGUCGUAGAUAAAAUAUUCAGUGCAAGAUAUACAUAUAAU GUUGAAAAACAAAGAUAUAAUAAUAAAUUUUCAUCCUCUAAUA AUUCUGUAUAUAAUGUUCAAAAAUUAAAAAAGGCUCUUUCAUA UCUUGAAGAUUAUUCUUUAAGAAAAGGAAUUUCUGAAAAAGA UUUUAAUCAUUAUUAUACUUUGAAAACUGGCCUCGAAGCUGAU AUAAAAAAAUUAACAGAAGAAAUAAAGAGUAGUGAAAACAAA AUUCUUGAAAAAAAUUUUAAAGGACUAACACAUUCAGCAAAUG GUUCCUUAGAAGUAUCUGAUAUUGUAAAAUUACAAGUACAAAA AGUUUUAUUAAUUAAAAAAAUAGAAGACUUAAGAAAGAUAGA AUUAUUUUUAAAAAAUGCACAACUAAAAGAUAGUAUUCAUGUA CCAAAUAUUUAUAAACCACAAAAUAAACCAGAACCAUAUUAUU UAAUUGUAUUAAAAAAAGAAGUAGAUAAAUUAAAAGAAUUUA UACCAAAAGUAAAAGACAUGUUAAAGAAAGAACAAGCUGUCUU AUCAAGUAUUACACAACCUUUAGUUGCAGCAAGCGAAACAACU GAAGAUGGGGGUCACUCCACACACACAUUAUCCCAAUCAGGAG AAACAGAAGUAACAGAAGAAACAGAAGAAACAGAAGAAACAGU AGGACACACAACAACGGUAACAAUAACAUUACCACCAACACAA CCAUCACCACCAAAAGAAGUAAAAGUUGUUGAAAAUUCAAUAG AACAUAAGAGUAAUGACAAUUCACAAGCCUUGACAAAAACAGU UUAUCUAAAGAAAUUAGAUGAAUUUUUAACUAAAUCAUAUAU AUGUCAUAAAUAUAUUUUAGUAUCAAACUCUAGUAUGGACCAA AAAUUAUUAGAGGUAUAUAAUCUUACUCCAGAAGAAGAAAAUG AAUUAAAAUCAUGUGAUCCAUUAGAUUUAUUAUUUAAUAUUCA AAAUAACAUACCUGCUAUGUAUUCAUUAUAUGAUAGUAUGAAC AAUGAUUUACAACAUCUCUUUUUUGAAUUAUAUCAAAAGGAAA UGAUUUAUUAUUUACAUAAACUAAAAGAGGAAAAUCACAUCAA AAAAUUAUUAGAGGAGCAAAAACAAAUAACUGGAACAUCAUCU ACAUCCAGUCCUGGAAAUACAACCGUAAAUACUGCUCAAUCCG CAACUCACAGUAAUUCCCAAAACCAACAAUCAAAUGCAUCCUC UACCAAUACCCAAAAUGGUGUAGCUGUAUCAUCUGGUCCUGCU GUAGUUGAAGAAAGUCAUGAUCCCUUAACAGUAUUGUCUAUUA GUAACGAUUUGAAAGGUAUUGUUAGUCUCUUAAAUCUUGGAAA UAAAACUAAAGUACCUAAUCCAUUAACCAUUUCUACAACAGAG AUGGAAAAAUUUUAUGAGAAUAUUUUAAAAAAUAAUGAUACC UAUUUUAAUGAUGAUAUCAAACAAUUCGUAAAAUCUAAUUCAA AAGUAAUUACAGGUUUGACCGAAACACAAAAAAAUGCAUUAAA UGAUGAAAUUAAAAAAUUAAAAGAUACUUUACAGUUAUCAUU UGAUUUAUAUAAUAAAUAUAAAUUAAAAUUAGAUAGAUUAUU UAAUAAGAAAAAAGAACUUGGCCAAGACAAAAUGCAAAUUAAA AAACUUACUUUAUUAAAAGAACAAUUAGAAUCAAAAUUGAAUU CACUUAAUAACCCACAUAAUGUAUUACAAAACUUUUCUGUUUU CUUUAACAAAAAAAAAGAAGCUGAAAUAGCAGAAACUGAAAAC ACAUUAGAAAACACAAAAAUAUUAUUGAAACAUUAUAAAGGAC UUGUUAAAUAUUAUAAUGGUGAAUCAUCUCCAUUAAAAACUUU AAGUGAAGUAUCAAUUCAAACAGAAGAUAAUUAUGCCAAUUUA GAAAAAUUUAGAGUAUUAAGUAAAAUAGAUGGAAAACUCAAU GAUAAUUUACAUUUAGGAAAGAAAAAAUUAUCUUUCUUAUCAA GUGGAUUACAUCAUUUAAUUACUGAAUUAAAAGAAGUAAUAA AAAAUAAAAAUUAUACAGGUAAUUCUCCAAGUGAAAAUAAUAA GAAAGUUAACGAAGCUUUAAAAUCUUACGAAAAUUUUCUCCCA GAAGCAAAAGUUACAACAGUUGUAACUCCACCUCAACCAGAUG UAACUCCAUCUCCAUUAUCUGUAAGGGUAAGUGGUAGUUCAGG AUCCACAAAAGAAGAAACACAAAUACCAACUUCAGGCUCUUUA UUAACAGAAUUACAACAAGUAGUACAAUUACAAAAUUAUGACG AAGAAGAUGAUUCCUUAGUUGUAUUACCCAUUUUUGGAGAAUC CGAAGAUAAUGACGAAUAUUUAGAUCAAGUAGUAACUGGAGAA GCAAUAUCUGUCACAAUGGAUAAUAUCCUCUCAGGAUUUGAAA AUGAAUAUGAUGUUAUAUAUUUAAAACCUUUAGCUGGAGUAU AUAGAAGCUUAAAAAAACAAAUUGAAAAAAACAUUUUUACAUU UAAUUUAAAUUUGAACGAUAUCUUAAAUUCACGUCUUAAGAAA CGAAAAUAUUUCUUAGAUGUAUUAGAAUCUGAUUUAAUGCAAU UUAAACAUAUAUCCUCAAAUGAAUACAUUAUUGAAGAUUCAUU UAAAUUAUUGAAUUCAGAACAAAAAAACACACUUUUAAAAAGU UACAAAUAUAUAAAAGAAUCAGUAGAAAAUGAUAUUAAAUUU GCACAGGAAGGUAUAAGUUAUUAUGAAAAGGUUUUAGCGAAA UAUAAGGAUGAUUUAGAAUCAAUUAAAAAAGUUAUCAAAGAA GAAAAGGAGAAGUUCCCAUCAUCACCACCAACAACACCUCCGU CACCAGCAAAAACAGACGAACAAAAGAAGGAAAGUAAGUUCCU UCCAUUUUUAACAAACAUUGAGACCUUAUACAAUAACUUAGUU AAUAAAAUUGACGAUUACUUAAUUAACUUAAAGGCAAAGAUUA ACGAUUGUAAUGUUGAAAAAGAUGAAGCACAUGUUAAAAUAAC UAAACUUAGUGAUUUAAAAGCAAUUGAUGACAAAAUAGAUCUU UUUAAAAACCCUUACGACUUCGAAGCAAUUAAAAAAUUGAUAA AUGAUGAUACGAAAAAAGAUAUGCUUGGCAAAUUACUUAGUAC AGGAUUAGUUCAAAAUUUUCCUAAUACAAUAAUAUCAAAAUUA AUUGAAGGAAAAUUCCAAGAUAUGUUAAACAUUUCACAACACC AAUGCGUAAAAAAACAAUGUCCAGAAAAUUCUGGAUGUUUCAG ACAUUUAGAUGAAAGAGAAGAAUGUAAAUGUUUAUUAAAUUA CAAACAAGAAGGUGAUAAAUGUGUUGAAAAUCCAAAUCCUACU UGUAACGAAAAUAAUGGUGGAUGUGAUGCAGAUGCCACAUGUA CCGAAGAAGAUUCAGGUAGCAGCAGAAAGAAAAUCACAUGUGA AUGUACUAAACCUGAUUCUUAUCCACUUUUCGAUGGUAUUUUC UGCAGUUCCUCUAACUUCUUAGGAAUAUCAUUCUUAUUAAUAC UCAUGUUAAUAUUAUACAGUUUCAUU AMA1_3D7_FL_ AUGAGAAAAUUAUACUGCGUAUUAUUAUUGAGCGCCUUUGAGU 11 DX UUACAUAUAUGAUAAACUUUGGAAGAGGACAGAAUUAUUGGG AACAUCCAUAUCAAAAUAGUGAUGUGUAUCGUCCAAUCAACGA ACAUAGGGAACAUCCAAAAGAAUACGAAUAUCCAUUACACCAG GAACAUACAUACCAACAAGAAGAUUCAGGAGAAGACGAAAAUA CAUUACAACACGCAUAUCCAAUAGACCACGAAGGUGCUGAACC CGCACCACAAGAACAAAACUUAUUCUCAAGCAUUGAAAUAGUC GAACGGUCGAAUUAUAUGGGUAAUCCGUGGACGGAAUACAUGG CAAAAUAUGAUAUUGAAGAAGUUCAUGGUUCAGGUAUAAGGG UAGAUUUAGGAGAGGAUGCUGAAGUAGCUGGAACUCAAUACAG ACUUCCAUCAGGUAAAUGUCCAGUAUUCGGUAAAGGUAUAAUU AUUGAGAAUUCAAAUACUACUUUUCUAACACCGGUAGCUACAG GAAAUCAAUAUCUAAAAGAUGGAGGGUUUGCGUUUCCUCCAAC AGAACCUCUUAUGUCACCAAUGACACUAGAUGAAAUGAGACAU UUUUAUAAGGAUAAUAAAUACGUAAAAAACCUCGACGAAUUGA CUUUAUGUUCAAGACAUGCAGGGAAUAUGAUUCCAGAUAAUGA UAAAAACUCAAAUUAUAAAUACCCGGCUGUUUAUGAUGAUAAA GAUAAAAAGUGUCAUAUCCUGUAUAUUGCAGCUCAAGAAAAUA AUGGUCCUAGAUAUUGUAAUAAAGACGAAAGUAAAAGAAACAG CAUGUUUUGUUUCAGACCAGCAAAAGACAUAAGCUUCCAAAAC UAUACUUACUUGUCUAAAAAUGUAGUUGAUAACUGGGAAAAAG UCUGCCCUAGAAAGAAUUUACAGAAUGCGAAAUUCGGACUAUG GGUCGAUGGUAACUGUGAAGAUAUACCACAUGUAAAUGAAUUU CCAGCAAUAGAUCUUUUCGAAUGCAAUAAACUGGUUUUUGAAU UGAGUGCCUCGGAUCAGCCAAAACAGUAUGAACAACAUUUAAC AGAUUACGAAAAAAUCAAAGAAGGUUUUAAAAAUAAAAAUGC UAGUAUGAUCAAGAGUGCUUUUCUUCCCACGGGUGCUUUUAAG GCAGAUCGUUAUAAAAGUCACGGUAAAGGAUAUAAUUGGGGUA ACUAUAACACAGAGACACAAAAGUGUGAAAUUUUUAAUGUUAA ACCAACAUGUCUCAUAAACAAUUCCUCAUACAUUGCUACCACU GCUUUAUCCCAUCCCAUCGAAGUAGAGAACAAUUUUCCUUGUA GCCUAUACAAAGAUGAAAUAAUGAAAGAAAUCGAGAGAGAAUC AAAACGAAUUAAAUUAAAUGAUAACGAUGACGAGGGGAACAAA AAAAUUAUAGCUCCACGCAUUUUUAUUUCAGACGAUAAAGACU CGUUAAAAUGCCCAUGUGACCCUGAAAUGGUCAGUAACAGUAC AUGCCGCUUCUUUGUAUGUAAAUGUGUAGAAAGGAGGGCAGAA GUAACAUCAAAUAACGAGGUUGUAGUAAAAGAAGAAUACAAAG AUGAGUACGCAGAUAUUCCCGAACACAAACCAACCUAUGAUAA GAUGAAAAUAAUAAUAGCAUCAUCAGCUGCAGUUGCUGUACUA GCGACGAUUUUAAUGGUUUAUCUUUAUAAAAGAAAGGGAAAU GCUGAAAAAUAUGAUAAAAUGGAUGAACCGCAAGAUUAUGGGA AAUCCAAUUCACGAAACGAUGAGAUGCUUGAUCCUGAGGCAUC UUUCUGGGGGGAAGAAAAAAGAGCAUCACAUACAACACCAGUA CUGAUGGAAAAACCAUACUAU PF3D7_1335900 | AUGAAUCAUCUUGGGAAUGUUAAAUAUUUAGUCAUUGUGUUU 12 Plasmodium UUGAUUUUCUUUGAUUUGUUUCUAGUUAAUGGUAGAGAUGUG falciparum 3D7 | CAAAACAAUAUAGUGGAUGAAAUAAAAUAUCGUGAAGAAGUA thrombospondin- UGUAAUGAUGAGGUAGAUCUUUACCUUCUAAUGGAUUGUUCUG related GAAGUAUACGUCGUCAUAAUUGGGUGAACCAUGCAGUACCUCU anonymous AGCUAUGAAAUUGAUACAACAAUUAAAUCUUAAUGAUAAUGCA protein (TRAP) | AUUCACUUAUAUGCUAGUGUUUUUUCAAACAAUGCAAGAGAAA CDS | UUAUUAGAUUACAUAGUGAUGCAUCUAAAAACAAAGAGAAGGC length = 1725 UUUAAUUAUUAUAAAGUCACUCUUAAGUACAAAUCUUCCAUAU GGUAAAACAAACUUAACUGAUGCACUGUUACAAGUAAGAAAAC AUUUAAAUGACCGAAUCAAUAGAGAGAAUGCUAAUCAAUUAGU UGUUAUAUUAACAGAUGGAAUUCCAGAUAGUAUUCAAGAUUCA UUAAAAGAAUCAAGAAAAUUAAGUGAUCGUGGUGUUAAAAUA GCUGUUUUUGGUAUUGGACAAGGUAUUAAUGUAGCUUUCAACA GAUUUCUUGUAGGUUGUCAUCCAUCAGAUGGUAAAUGUAACUU GUAUGCUGAUUCUGCAUGGGAAAAUGUAAAAAAUGUUAUCGGA CCCUUUAUGAAGGCUGUUUGUGUUGAAGUAGAAAAAACAGCAA GUUGUGGUGUUUGGGACGAAUGGUCUCCAUGUAGUGUAACUUG UGGUAAAGGUACCAGGUCAAGAAAAAGAGAAAUCUUACACGAA GGAUGUACAAGUGAAUUACAAGAACAAUGUGAAGAAGAAAGA UGUCUUCCAAAACGGGAACCAUUAGAUGUUCCAGAUGAACCCG AAGAUGAUCAACCUAGACCAAGAGGAGAUAAUUUUGCUGUCGA AAAACCAAACGAAAAUAUAAUAGAUAAUAAUCCACAAGAACCU UCACCAAAUCCAGAAGAAGGAAAGGGUGAAAAUCCAAACGGAU UUGAUUUAGAUGAAAAUCCAGAAAAUCCACCAAAUCCACCAAA UCCACCAAAUCCACCAAAUCCACCAAAUCCACCAAAUCCAGAUA UUCCUGAACAAGAACCAAAUAUACCUGAAGAUUCAGAAAAAGA AGUACCUUCUGAUGUUCCAAAAAAUCCAGAAGACGAUCGAGAA GAAAACUUUGAUAUUCCAAAGAAACCCGAAAAUAAGCACGAUA AUCAAAAUAAUUUACCAAAUGAUAAAAGUGAUAGAUAUAUUCC AUAUUCACCAUUAUCUCCAAAAGUUUUGGAUAAUGAAAGGAAA CAAAGUGACCCCCAAAGUCAAGAUAAUAAUGGAAAUAGGCACG UACCUAAUAGUGAAGAUAGAGAAACACGUCCACAUGGUAGAAA UAAUGAAAAUAGAUCAUACAAUAGAAAACAUAACAAUACUCCA AAACAUCCUGAAAGGGAAGAACAUGAAAAGCCAGAUAAUAAUA AAAAAAAAGCAGGAUCAGAUAAUAAAUAUAAAAUUGCAGGUG GAAUAGCUGGAGGAUUAGCUUUACUCGCAUGUGCUGGACUUGC UUAUAAAUUCGUAGUACCAGGAGCAGCAACACCCUAUGCCGGA GAACCUGCACCUUUUGAUGAAACAUUAGGUGAAGAAGAUAAAG AUUUGGACGAACCUGAACAAUUCAGAUUACCUGAAGAAAACGA GUGGAAUUAA

TABLE 2 Plasmodium Amino Acid Sequences Description Sequences SEQ ID NO: PF3D7_1335900 | MNHLGNVKYLVIVFLIFFDLFLVNGRDVQNNIVDEIKYREEVCNDEV 13 Plasmodium DLYLLMDCSGSIRRHNWVNHAVPLAMKLIQQLNLNDNAIHLYASINV falciparum 3D7 | FSNNAREIIRLHSDASKNKEKALIIIKSLLSTNLPYGKTNLTDALLQ thrombospondin- VRKHLNDRINRENANQLVVILTDGIPDSIQDSLKESRKLSDRGVKIA related VFGIGQGINVAFNRFLVGCHPSDGKCNLYADSAWENVKNVIGPFMKA anonymous VCVEVEKTASCGVWDEWSPCSINVTCGKGTRSRKREILHEGCTSELQ protein (TRAP) | EQCEEEVRCLPKREPLDVPDEPEDDQPRPRGDNFAVEKPNENIIDNN protein | PQEPSPNPEEGKGENPNGFDLDENPENPPNPPNPPNPPNPPNPPNPD length = 574 IPEQEPNIPEDSEKEVPSDVPKNPEDDREENFDIPKKPENKHDNQNN LPNDKSDRYIPYSPLSPKVLDNERKQSDPQSQDNNGNRHVPNSEDRE TRPHGRNNENRSYNRKHNNTPKHPEREEHEKPDNNKKKAGSDNKYKI AGGIAGGLALLACAGLAYKFVVPGAATPYAGEPAPFDETLGEEDKDL DEPEQFRLPEENEWN apical membrane MRKLYCVLLLSAFEFTYMINFGRGQNYWEHPYQNSDVYRPINEHREH 14 antigen 1, AMA1 PKEYEYPLHQEHTYQQEDSGEDENTLQHAYPIDHEGAEPAPQEQNLF [Plasmodium SSIEIVERSNYMGNPWTEYMAKYDIEEVHGSGIRVDLGEDAEVAGTQ falciparum 3D7] YRLPSGKCPVFGKGIIIENSNTTFLTPVATGNQYLKDGGFAFPPTEP LMSPMTLDEMRHFYKDNKYVKNLDELTLCSRHAGNMIPDNDKNSNYK YPAVYDDKDKKCHILYIAAQENNGPRYCNKDESKRNSMFCFRPAKDI SFQNYTYLSKNVVDNWEKVCPRKNLQNAKFGLWVDGNCEDIPHVNEF PAIDLFECNKLVFELSASDQPKQYEQHLTDYEKIKEGFKNKNASMIK SAFLPTGAFKADRYKSHGKGYNWGNYNTETQKCEIFNVKPTCLINNS SYIATTALSHPIEVENNFPCSLYKDEIMKEIERESKRIKLNDNDDEG NKKIIAPRIFISDDKDSLKCPCDPEMVSNSTCRFFVCKCVERRAEVT SNNEVVVKEEYKDEYADIPEHKPTYDKMKIIIASSAAVAVLATILMV YLYKRKGNAEKYDKMDEPQDYGKSNSRNDEMLDPEASFWGEEKRASH TTPVLMEKPYY merozoite surface MKIIFFLCSFLFFIINTQCVTHESYQELVKKLEALEDAVLTGYSLFQ 15 protein 1 KEKMVLNEEEVITTKGASAQSGASAQSGASAQSGASAQSGASAQSGA precursor SAQSGTSGPSGPSGTSPSSRSNTLPRSNTSSGASPPADASDSDAKSY [Plasmodium ADLKHRVRNYLFTIKELKYPELFDLTNHMLTLCDNIHGFKYLIDGYE falciparum 3D7] EINELLYKLNFYFDLLRAKLNDVCANDYCQIPFNLKIRANELDVLKK LVFGYRKPLDNIKDNVGKMEDYIKKNKTTIANINELIEGSKKTIDQN KNADNEEGKKKLYQAQYDLSIYNKQLEEAHNLISINVLEKRIDTLKK NENIKKLLDKINEIKNPPPANSGNTPNTLLDKNKKIEEHEEKIKEIA KTIKFNIDSLFTDPLELEYYLREKNKKVDVTPKSQDPTKSINVQIPK VPYPNGIVYPLPLTDIHNSLAADNDKNSYGDLMNPHTKEKINEKIIT DNKERKIFINNIKKKIDLEEKNINHTKEQNKKLLEDYEKSKKDYEEL LEKFYEMKFNNNFNKDVVDKIFSARYTYNVEKQRYNNKFSSSNNSIN VYNVQKLKKALSYLEDYSLRKGISEKDFNHYYTLKTGLEADIKKLTE EIKSSENKILEKNFKGLTHSANGSLEVSDIVKLQVQKVLLIKKIEDL RKIELFLKNAQLKDSIHVPNIYKPQNKPEPYYLIVLKKEVDKLKEFI PKVKDMLKKEQAVLSSITQPLVAASETTEDGGHSTHTLSQSGETEVT EETEETEETVGHTTTVTITLPPTQPSPPKEVKVVENSIEHKSNDNSQ ALTKTVYLKKLDEFLTKSYICHKYILVSNSSMDQKLLEVYNLTPEEE VNELKSCDPLDLLFNIQNNIPAMYSLYDSMNNDLQHLFFELYQKEMI YYLHKLKEENHIKKLLEEQKQITGTSSTSSPGNTTVNTAQSATHSNS QNQQSNASSTNTQNGVAVSSGPAVVEESHDPLTVLSISNDLKGIVSL LNLGNKTKVPNPLTISTTEMEKFYENILKNNDTYFNDDIKQFVKSNS KVITGLTETQKNALNDEIKKLKDTLQLSFDLYNKYKLKLDRLFNKKK ELGQDKMQIKKLTLLKEQLESKLNSLNNPHNVLQNFSINVFFNKKKE AEIAETENTLENTKILLKHYKGLVKYYNGESSPLKTLSEVSIQTEDN YANLEKFRVLSKIDGKLNDNLHLGKKKLSFLSSGLHHLITELKEVIK NKNYTGNSPSENNKKVNEALKSYENFLPEAKVTTVVTPPQPDVTPSP LSINVRVSGSSGSTKEETQIPTSGSLLTELQQVVQLQNYDEEDDSLV VLPIFGESEDNDEYLDQVVTGEAISINVTMDNILSGFENEYDVIYLK PLAGVYRSLKKQIEKNIFTFNLNLNDILNSRLKKRKYFLDVLESDLM QFKHISSNEYIIEDSFKLLNSEQKNTLLKSYKYIKESINVENDIKFA QEGISYYEKVLAKYKDDLESIKKVIKEEKEKFPSSPPTTPPSPAKTD EQKKESKFLPFLTNIETLYNNLVNKIDDYLINLKAKINDCNVEKDEA HVKITKLSDLKAIDDKIDLFKNPYDFEAIKKLINDDTKKDMLGKLLS TGLVQNFPNTIISKLIEGKFQDMLNISQHQCVKKQCPENSGCFRHLD EREECKCLLNYKQEGDKCVENPNPTCNENNGGCDADATCTEEDSGSS RKKITCECTKPDSYPLFDGIFCSSSNFLGISFLLILMLILYSFI liver stage antigen MKHILYISFYFILVNLLIFQINGKILKKSENDEIIKSNLRSGSSNSL 16 1 [Plasmodium NQIYEEKHEKKHALSHNSYDKTKNNENHKFFDKDKEVSISNLKNVSQ reichenowi] TNVKNALRNFGVSENIFLQENKLGEEGKLINHITNDDENKEKYIKGE EEVNREEDPEEKAARERQEAEEKAARERQEAEEKAAREKQEAEEKAA RERQVAEKAAREKQEAEEKAAREKQEAEEKAAREREESPKRALAEQR AAFERIDSMKQKLEGGKEHGDVLAEDLYGRLEIPVIELPSENEGGYY IQHQSSLPQDNRGNSRDSKEISIIEKTNRESITTNVEGRRDIHKGHL EEKKDVSIKPEQKEDKSADVQNHALETVNILDVKDFQISEYEDEISA EYDDSLIDEEEVDDEDLDQFKPIVQYDNFQDEENIGIYKELEDLIEK NENLDDLDEGIEKSSEELSEEKKKKGKKYEKTKDTNFKPNDKSLYDE HIKKYKNVKQINKEKEKFIKSLFHIFDGDNEILQIVDELSEDITKYF MKL circumsporozoite MMRKLAILSINVSSFLFVEALFQEYQCYGSSSNTRVLNELNYDNAGT  17 (CS) protein NLYNELEMNYYGKQENWYSLKKNSRSLGENDDGNNEDNEKLRKPKHK [Plasmodium KLKQPADGNPDPNANPNVDPNANPNVDPNANPNVDPNANPNANPNAN falciparum 3D7] PNANPNANPNANPNANPNANPNANPNANPNANPNANPNANPNANPNA NPNANPNANPNVDPNANPNANPNANPNANPNANPNANPNANPNANPN ANPNANPNANPNANPNANPNANPNANPNANPNANPNANPNKNNQGNG QGHNMPNDPNRNVDENANANSAVKNNNNEEPSDKHIKEYLNKIQNSL STEWSPCSINVTCGNGIQVRIKPGSANKPKDELDYANDIEKKICKME KCSSINVFNVVNSSIGLIMVLSFLFLN

TABLE 3 Plasmodium NCBI Accession Numbers (Amino Acid Sequences) Protein Name GenBank Accession AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAM85704.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAM85539.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AHI17058.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAM85463.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AHI16999.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAM85538.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACB87847.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AHI17042.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACB87818.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACB87776.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAM85414.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28650.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAM85410.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACB87856.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28916.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28914.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACR49683.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACB87823.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAM85642.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAM85598.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAM85451.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACR49654.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28923.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACR49728.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACR49672.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACB87806.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28573.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACA24168.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACB87765.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACB87843.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAM85489.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAM85383.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28825.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACR49680.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACR49668.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAN59369.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAM85460.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAM85443.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28834.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28579.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACR49756.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACB87811.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACB87763.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAM85525.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28604.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAM85651.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAM85430.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28822.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACB87817.1 AMA1 apical membrane antigen 1, AMA1 [Plasmodium falciparum 3D7] XP_001348015.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum FCH/4] ETW28666.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAM85388.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28551.1 AMA1 apical membrane antigen-1 [Plasmodium falciparum] AAC47104.1 AMA1 apical membrane antigen-1 [Plasmodium falciparum] AAG50121.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACA24148.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAN59390.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAM85527.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28590.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28523.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACR49659.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAM85518.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAM85401.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28892.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28835.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28666.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28610.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACB87841.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28663.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACR49754.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACB87786.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAM85680.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28660.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACB87787.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28864.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28576.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACR49702.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28924.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28732.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAM85529.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28987.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28645.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28547.1 AMA1 RecName: Full = Apical membrane antigen 1; AltName: Full = Merozoite surface antigen; Flags: Precursor P22621.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACR49695.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AHI17024.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACB87801.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28736.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28661.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28669.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAM85445.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAM85373.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACR49655.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28826.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAM85366.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28755.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] ACB87802.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AHI17004.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] BAM85450.1 AMA1 apical membrane antigen 1 [Plasmodium falciparum] AFM28965.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] ACB55391.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] AKC42164.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68759.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] ACB55341.1 AMA1 merozoite receptor PK66 [Plasmodium vivax Brazil I] KMZ86684.1 AMA1 merozoite receptor PK66 [Plasmodium vivax North Korean] KMZ99594.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] ACB55347.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] ACB55385.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] ACB55371.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68738.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] AKC42165.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] ACB55389.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] AKC42198.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] ACB55320.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68744.1 AMA1 apical merozoite protein 1 [Plasmodium vivax] ACB42437.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] ACB55357.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] ABM63524.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] AKC42208.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68762.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] AKC42207.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68808.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68841.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68809.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68799.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68778.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] ACB55354.1 AMA1 apical merozoite protein 1 [Plasmodium vivax] ACB42433.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] ACB55334.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68789.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68769.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] ACB55383.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68736.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] ACB55322.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] AKC42213.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] AKC42203.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68774.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68754.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68782.1 AMA1 merozoite receptor PK66 [Plasmodium vivax Mauritania I] KMZ93134.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68766.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] AKC42211.1 AMA1 apical membrane antigen [Plasmodium vivax] AKQ33081.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68827.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] AKC42170.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68849.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68753.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] ABM63521.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68850.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68795.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68739.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] ACB55332.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] AKC42163.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68790.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68751.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68763.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68746.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] ACB55378.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] AKC42218.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68889.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68829.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68771.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68726.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] ABM63525.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68775.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68750.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] ACB55319.1 AMA1 apcial membrane antigen I [Plasmodium vivax] AAA29480.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68807.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68779.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68756.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68752.1 AMA1 apcial membrane antigen I [Plasmodium vivax] AAA29479.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] AKC42169.1 AMA1 apical merozoite antigen 1 [Plasmodium vivax Sal-1] XP_001615447.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] AKC42185.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] AKC42166.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68881.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68802.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68773.1 AMA1 apical membrane antigen-1 [Plasmodium vivax] BAH96576.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68837.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68830.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68761.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] ACB55369.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] AKC42184.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68742.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68730.1 AMA1 apical membrane antigen 1 [Plasmodium vivax] AKC42167.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68880.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68791.1 AMA1 apical merozoite protein 1 [Plasmodium vivax] ACB42435.1 AMA1 apical membrane antigen [Plasmodium vivax] AKQ33082.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68820.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68804.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68737.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68733.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68935.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68871.1 AMA1 apical membrane protein-1 [Plasmodium vivax] ACY68825.1 CSP RecName: Full = Circumsporozoite protein; Short = CS; Flags: Precursor P02893.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84780.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84770.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84757.1 CSP circumsporozoite protein [Plasmodium falciparum] AAN87606.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84998.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84921.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84784.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84763.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84804.1 CSP circumsporozoite protein [Plasmodium falciparum] AAN87603.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84759.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84963.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84807.1 CSP circumsporozoite protein [Plasmodium falciparum] ACO49330.1 CSP circumsporozoite protein [Plasmodium falciparum] BAD08404.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84779.1 CSP circumsporozoite protein [Plasmodium falciparum] AAN87586.1 CSP circumsporozoite protein [Plasmodium falciparum] AGR53780.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM85040.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84768.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84753.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84923.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84806.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84797.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84750.1 CSP circumsporozoite protein [Plasmodium falciparum] AAA29550.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84801.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84775.1 CSP circumsporozoite protein [Plasmodium falciparum] AAN87601.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84935.1 CSP circumsporozoite protein [Plasmodium falciparum] BAD08405.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84975.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84971.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84760.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84792.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84781.1 CSP circumsporozoite protein [Plasmodium falciparum] AAN87598.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM85004.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM85007.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84962.1 CSP circumsporozoite protein [Plasmodium falciparum] AAN87611.1 CSP circumsporozoite protein [Plasmodium falciparum] BAD08407.1 CSP circumsporozoite protein [Plasmodium falciparum] AAW59565.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84927.1 CSP circumsporozoite protein [Plasmodium falciparum] AAA63422.1 CSP circumsporozoite protein [Plasmodium falciparum] ACO49339.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84917.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84773.1 CSP circumsporozoite protein [Plasmodium falciparum] BAD08408.1 CSP circumsporozoite protein [Plasmodium falciparum] AAN87589.1 CSP circumsporozoite protein [Plasmodium falciparum] ACO49375.1 CSP circumsporozoite protein [Plasmodium falciparum] AAA29547.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84946.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84764.1 CSP circumsporozoite protein [Plasmodium falciparum] ACO49332.1 CSP circumsporozoite protein [Plasmodium falciparum] BAD73949.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84820.1 CSP circumsporozoite protein [Plasmodium falciparum] ACO49474.1 CSP circumsporozoite protein [Plasmodium falciparum] ACO49328.1 CSP circumsporozoite protein [Plasmodium falciparum] ACO49327.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84938.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84765.1 CSP circumsporozoite protein [Plasmodium falciparum] ACO49410.1 CSP circumsporozoite protein [Plasmodium falciparum] BAD08409.1 CSP circumsporozoite protein [Plasmodium falciparum] ACO49331.1 CSP RecName: Full = Circumsporozoite protein; Short = CS; Flags: Precursor P13814.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84758.1 CSP circumsporozoite protein [Plasmodium falciparum] ACO49492.1 CSP circumsporozoite protein [Plasmodium falciparum] ACO49333.1 CSP circumsporozoite protein [Plasmodium falciparum] AAN87612.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84936.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84856.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84798.1 CSP circumsporozoite protein [Plasmodium falciparum] BAD73952.1 CSP circumsporozoite protein [Plasmodium falciparum] AAN87590.1 CSP circumsporozoite protein [Plasmodium falciparum] AAA29544.1 CSP circumsporozoite protein [Plasmodium falciparum] AAN87591.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84896.1 CSP circumsporozoite protein [Plasmodium falciparum] ACO49368.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84895.1 CSP circumsporozoite protein [Plasmodium falciparum] BAD73951.1 CSP circumsporozoite protein [Plasmodium falciparum] AAA29543.1 CSP circumsporozoite protein [Plasmodium falciparum] AAN87614.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84918.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84893.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84800.1 CSP circumsporozoite protein [Plasmodium falciparum] ACO49325.1 CSP circumsporozoite protein [Plasmodium falciparum] AAA29552.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84953.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84813.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84790.1 CSP circumsporozoite protein [Plasmodium falciparum] ACO49384.1 CSP circumsporozoite protein [Plasmodium falciparum] AAN87609.1 CSP circumsporozoite protein [Plasmodium falciparum] AAA29574.1 CSP circumsporozoite protein [Plasmodium falciparum] ACO49409.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84989.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84930.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84805.1 CSP circumsporozoite protein [Plasmodium falciparum] BAM84756.1 CSP RecName: Full = Circumsporozoite protein; Short = CS; Flags: Precursor P08677.2 PV1H14150_P [Plasmodium vivax] AAF99476.1 CSP circumsporozoite protein [Plasmodium vivax] ACB38256.1 CSP circumsporozoite protein [Plasmodium vivax] AGN05254.1 CSP circumsporozoite protein [Plasmodium vivax] ADB92541.1 CSP RecName: Full = Circumsporozoite protein; Short = CS; Flags: Precursor Q03110.1 CSP circumsporozoite protein [Plasmodium vivax] ADB92554.1 CSP circumsporozoite protein precursor [Plasmodium vivax Sal-1] XP_001613068.1 CSP circumsporozoite protein [Plasmodium vivax] BAO10686.1 CSP circumsporozoite protein [Plasmodium vivax] AAA18615.1 CSP circumsporozoite protein [Plasmodium vivax] BAO10687.1 CSP circumsporozoite protein [Plasmodium vivax] BAO10684.1 CSP circumsporozoite protein [Plasmodium vivax] ACB38257.1 CSP circumsporozoite protein [Plasmodium vivax] BAO10683.1 CSP circumsporozoite protein [Plasmodium vivax] BAO10674.1 CSP circumsporozoite protein [Plasmodium vivax] AAA29535.1 CSP circumsporozoite protein [Plasmodium vivax] BAO10694.1 CSP circumsporozoite protein [Plasmodium vivax] BAO10677.1 CSP circumsporozoite protein [Plasmodium vivax] ACB38264.1 CSP circumsporozoite protein [Plasmodium vivax] AGN05250.1 CSP circumsporozoite protein [Plasmodium vivax] AGN05243.1 CSP circumsporozoite protein [Plasmodium vivax] AGN05261.1 CSP circumsporozoite protein [Plasmodium vivax] BAO10680.1 CSP circumsporozoite protein [Plasmodium vivax] AGN05251.1 CSP circumsporozoite protein [Plasmodium vivax] BAO10696.1 CSP circumsporozoite protein [Plasmodium vivax] AGN05247.1 CSP circumsporozoite protein [Plasmodium vivax] BAO10693.1 CSP circumsporozoite protein [Plasmodium vivax] BAO10673.1 CSP circumsporozoite protein [Plasmodium vivax] BAO10679.1 CSP circumsporozoite protein [Plasmodium vivax] BAO10688.1 CSP circumsporozoite protein [Plasmodium vivax] BAO10685.1 CSP circumsporozoite protein [Plasmodium vivax] BAO10678.1 CSP circumsporozoite protein [Plasmodium vivax] AGN05262.1 CSP circumsporozoite protein [Plasmodium vivax] BAO10692.1 CSP circumsporozoite protein [Plasmodium vivax] BAO10691.1 CSP circumsporozoite protein [Plasmodium vivax] AGN05249.1 CSP circumsporozoite protein [Plasmodium vivax] AGN05253.1 CSP circumsporozoite protein [Plasmodium vivax] AAZ81578.1 CSP circumsporozoite protein [Plasmodium vivax] AGN05246.1 CSP circumsporozoite protein [Plasmodium vivax] AGN05273.2 CSP circumsporozoite protein [Plasmodium vivax] AGN05232.1 CSP circumsporozoite protein [Plasmodium vivax] AGN05237.1 CSP circumsporozoite protein [Plasmodium vivax] AFI80543.1 CSP circumsporozoite protein [Plasmodium vivax] AFI80544.1 CSP circumsporozoite protein [Plasmodium vivax] AFI80542.1 CSP circumsporozoite protein [Plasmodium vivax] AGN05244.1 CSP circumsporozoite protein [Plasmodium vivax] AFI80541.1 CSP circumsporozoite protein [Plasmodium vivax] BAO10695.1 CSP circumsporozoite protein [Plasmodium vivax] AGN05245.1 CSP circumsporozoite protein [Plasmodium vivax] AHL69651.1 CSP circumsporozoite protein [Plasmodium vivax] AGN05258.1 CSP circumsporozoite protein [Plasmodium vivax] ACB38262.1 CSP circumsporozoite protein [Plasmodium vivax] ACB38260.1 CSP circumsporozoite protein [Plasmodium vivax] AAC46505.1 CSP circumsporozoite protein [Plasmodium vivax] ACB38259.1 CSP circumsporozoite protein [Plasmodium vivax] AFI80540.1 CSP RecName: Full = Circumsporozoite protein; Short = CS P13826.1 CSP circumsporozoite protein [Plasmodium vivax] AAC46502.1 CSP circumsporozoite protein [Plasmodium vivax] ACB38258.1 CSP CSP [Plasmodium vivax] ABJ52967.1 CSP circumsporozoite protein [Plasmodium vivax] AAC46504.1 CSP circumsporozoite protein [Plasmodium vivax] AAG53720.2 CSP circumsporozoite protein [Plasmodium vivax] ACN69811.1 CSP CSP [Plasmodium vivax] ABJ52997.1 CSP circumsporozoite protein [Plasmodium vivax] AAL30430.1 CSP circumsporozoite protein [Plasmodium vivax] AAN63613.1 CSP circumsporozoite protein variant VK210 [synthetic construct] AHC98629.1 CSP circumsporozoite protein [Plasmodium vivax] AAZ81579.1 CSP CSP [Plasmodium vivax] ABJ53007.1 CSP circumsporozoite protein [Plasmodium vivax] AAC46500.1 CSP CSP [Plasmodium vivax] ABJ53001.1 CSP circumsporozoite protein [Plasmodium vivax] AAC46501.1 CSP circumsporozoite protein [Plasmodium vivax] AAZ81581.1 CSP circumsporozoite protein [Plasmodium vivax] ABI55078.1 CSP circumsporozoite protein [Plasmodium vivax] ABI55049.1 CSP circumsporozoite protein [Plasmodium vivax] AGK92816.1 CSP circumsporozoite protein [Plasmodium vivax] ABI55063.1 CSP circumsporozoite protein [Plasmodium vivax] ABI55053.1 CSP circumsporozoite protein [Plasmodium vivax] ABI55054.1 CSP circumsporozoite protein [Plasmodium vivax] ABI55067.1 CSP circumsporozoite protein [Plasmodium vivax] AAZ81577.1 CSP circumsporozoite protein [Plasmodium vivax] AAZ81580.1 CSP circumsporozoite protein [Plasmodium vivax] AAZ81583.1 CSP circumsporozoite protein [Plasmodium vivax] AAZ81582.1 CSP CSP [Plasmodium vivax] AAV80840.1 CSP circumsporozoite protein [Plasmodium vivax] ACN69845.1 CSP circumsporozoite protein [Plasmodium vivax] AAZ81587.1 CSP circumsporozoite protein [Plasmodium vivax] AAZ81584.1 CSP circumsporozoite protein [Plasmodium vivax] AFJ04722.1 CSP circumsporozoite protein [Plasmodium vivax] ABI55047.1 CSP circumsporozoite protein [Plasmodium vivax] AAZ81585.1 CSP circumsporozoite protein [Plasmodium vivax] AAC46499.1 CSP CSP [Plasmodium vivax] ABJ52988.1 CSP circumsporozoite protein [Plasmodium vivax] ACN69851.1 CSP circumsporozoite protein [Plasmodium vivax] AAA29534.1 CSP circumsporozoite protein [Plasmodium vivax] ACN69850.1 CSP CSP [Plasmodium vivax] ABJ53002.1 CSP circumsporozoite protein [Plasmodium vivax] ACN69873.1 CSP circumsporozoite protein [Plasmodium vivax] ACN69801.1 CSP circumsporozoite protein [Plasmodium vivax] ACN69855.1 CSP RecName: Full = Circumsporozoite protein; Short = CS; Flags: Precursor P13815.1 CSP circumsporozoite [Plasmodium Malariae] CAA04809.1 CSP circumsporozoite [Plasmodium Malariae] CAA04812.1 CSP circumsporozoite protein [Plasmodium Malariae] CAA05616.1 CSP circumsporozoite protein [Plasmodium Malariae] CAA05620.1 CSP circumsporozoite protein [Plasmodium Malariae] CAA05623.1 CSP circumsporozoite protein [Plasmodium Malariae] AAA18618.1 CSP circumsporozoite protein [Plasmodium Malariae] CAA05622.1 CSP circumsporozoite protein [Plasmodium Malariae] CAA05624.1 CSP circumsporozoite protein [Plasmodium Malariae] CAA05618.1 CSP circumsporozoite protein [Plasmodium Malariae] CAA05617.1 CSP circumsporozoite [Plasmodium Malariae] CAA04811.1 EMP1 EMP1 [Plasmodium falciparum] AEA03008.1 EMP1 erythrocyte membrane protein 1 [Plasmodium falciparum RAJ116] KNC35020.1 EMP1 EMP1 [Plasmodium falciparum] AFJ66677.1 EMP1 erythrocyte membrane protein 1 [Plasmodium falciparum] AAQ73923.1 EMP1 pfEMP1 [Plasmodium falciparum HB3] KOB63865.1 EMP1 erythrocyte membrane protein 1, EMP1 [Plasmodium reichenowi] XP_012760486.1 EMP1 erythrocyte membrane protein 1, EMP1 [Plasmodium reichenowi] XP_012760381.1 EMP1 erythrocyte membrane protein 1, EMP1 [Plasmodium reichenowi] XP_012760689.1 EMP1 erythrocyte membrane protein 1, PfEMP1 [Plasmodium falciparum 3D7] XP_001349740.1 EMP1 erythrocyte membrane protein 1, EMP1 [Plasmodium reichenowi] XP_012762123.1 EMP1 PfEMP1 [Plasmodium falciparum Dd2] KOB85186.1 EMP1 EMP1 [Plasmodium falciparum] AFJ66673.1 EMP1 erythrocyte membrane protein 1, EMP1 [Plasmodium reichenowi] XP_012763428.1 EMP1 erythrocyte membrane protein 1, EMP1 [Plasmodium reichenowi] XP_012760734.1 EMP1 EMP1 [Plasmodium falciparum] AFJ66666.1 EMP1 EMP1 [Plasmodium falciparum] AFJ66671.1 EMP1 PfEMP1 [Plasmodium falciparum Dd2] KOB84755.1 EMP1 erythrocyte membrane protein 1 [Plasmodium falciparum] AJD77408.1 EMP1 EMP1 [Plasmodium falciparum] AFJ66669.1 EMP1 erythrocyte membrane protein 1 [Plasmodium falciparum] AAQ73925.1 EMP1 erythrocyte membrane protein 1 [Plasmodium falciparum IGH-CR14] KNG75236.1 EMP1 erythrocyte membrane protein 1 [Plasmodium falciparum IGH-CR14] KNG77255.1 EMP1 variant-specific surface protein 1 [Plasmodium falciparum IGH-CR14] KNG75273.1 EMP1 erythrocyte membrane protein 1 [Plasmodium falciparum IGH-CR14] KNG74414.1 EMP1 erythrocyte membrane protein 1, PfEMP1 [Plasmodium falciparum 3D7] XP_001347692.1 EMP1 erythrocyte membrane protein 1 [Plasmodium falciparum IGH-CR14] KNG75311.1 EMP1 erythrocyte membrane protein 1 [Plasmodium falciparum IGH-CR14] KNG74203.1 EMP1 erythrocyte membrane protein 1 [Plasmodium falciparum] AAO67411.1 EMP1 erythrocyte membrane protein 1, EMP1 [Plasmodium reichenowi] XP_012760271.1 EMP1 erythrocyte membrane protein 1, EMP1 [Plasmodium reichenowi] XP_012761099.1 EMP1 erythrocyte membrane protein 1 [Plasmodium falciparum] AAQ73928.1 EMP1 erythrocyte membrane protein 1 var IT-ICAM [Plasmodium falciparum] AAS89259.1 EMP1 erythrocyte membrane protein 1, EMP1 [Plasmodium reichenowi] XP_012760435.1 EMP1 erythrocyte membrane protein 1, PfEMP1 [Plasmodium falciparum 3D7] XP_001351321.1 EMP1 EMP1 [Plasmodium falciparum] AFJ66667.1 EMP1 erythrocyte membrane protein 1 [Plasmodium falciparum] ABM88768.1 EMP1 erythrocyte membrane protein 1 [Plasmodium falciparum] AAL12845.1 EMP1 erythrocyte membrane protein 1 [Plasmodium falciparum] ABM88752.1 EMP1 variant-specific surface protein [Plasmodium falciparum] AAA75396.1 EMP1 erythrocyte membrane protein 1 [Plasmodium falciparum IGH-CR14] KNG74927.1 EMP1 PfEMP1 [Plasmodium falciparum] CAD20868.1 EMP1 erythrocyte membrane protein 1 [Plasmodium falciparum] AAM55194.1 EMP1 erythrocyte membrane protein 1 var IT-ICAM [Plasmodium falciparum IGH- KNG78240.1 CR14] EMP1 erythrocyte membrane protein 1 [Plasmodium falciparum] AJD77405.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAF62287.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84441.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84452.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84429.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84458.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84459.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAF62286.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84699.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84463.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84398.1 MSP1 merozoite surface protein 1 [Plasmodium falciparum] BAD08401.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84657.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84628.1 MSP1 merozoite surface protein 1 [Plasmodium falciparum] BAD08403.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84485.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84658.1 MSP1 merozoite surface protein 1 [Plasmodium falciparum] BAD08402.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84418.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84490.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAF62272.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84445.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84474.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84419.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84446.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84462.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84404.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84438.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84454.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84580.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84395.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84536.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum IGH-CR14] KNG75390.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84440.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84641.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84447.1 MSP1 merozoite surface protein 1 [Plasmodium falciparum] BAD08400.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84532.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84630.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAF62269.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84564.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84466.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84436.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84610.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84585.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAF62282.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84409.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84590.1 MSP1 merozoite surface protein 1 [Plasmodium falciparum FCH/4] ETW30902.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84467.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84684.1 MSP1 RecName: Full = Merozoite surface protein 1; AltName: Full = Merozoite P19598.2 surface antigens; AltName: Full = PMMSA; AltName: Full = p190; Flags: Precursor MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84468.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84478.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84633.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84556.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84504.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84456.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84475.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAF62283.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84685.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84553.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84547.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84403.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84555.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84477.1 MSP1 merozoite surface protein 1 [Plasmodium falciparum] BAN59439.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84618.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84539.1 MSP1 merozoite surface protein 1 [Plasmodium falciparum] BAN59436.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84603.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84714.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84655.1 MSP1 merozoite surface protein 1 [Plasmodium falciparum] BAD08399.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAF62285.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAF62275.1 MSP1 merozoite surface protein 1 [Plasmodium falciparum MaliPS096_E11] ETW49422.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84683.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84559.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84576.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84698.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84602.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAF62274.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84578.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84650.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84629.1 MSP1 merozoite surface protein 1 precursor [Plasmodium falciparum] AAC72884.1 MSP1 major merozoite surface antigen [Plasmodium falciparum] AAF27526.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84587.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAF62278.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84476.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84414.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84588.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84577.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84505.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84443.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84613.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84507.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84579.1 MSP1 merozoite surface protein-1 [Plasmodium falciparum] BAM84483.1 MSP1 merozoite surface protein 1 [Plasmodium falciparum] BAD08398.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] BAJ41320.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] AGM38068.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] AAN86238.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48827.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] AAN86218.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48832.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48583.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48786.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] AAN86208.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48837.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48548.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] AAN86243.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48567.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48797.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48789.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48581.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48812.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48646.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48561.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48795.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48785.1 MSP1 merozoite surface antigen 1 [Plasmodium vivax] AAA63427.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] AAN86210.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48554.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48586.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48577.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] ADN22980.1 MSP1 hypothetical protein PVNG_06384 [Plasmodium vivax North Korean] KNA00298.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48545.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] AAN86224.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] AAN86240.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] AAN86217.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48584.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] BAJ41310.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48834.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] AAN86246.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] ADN22981.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] ADN22979.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48558.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48550.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] AAN86231.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48841.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48839.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48818.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48564.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] AAN86242.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48810.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48568.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48559.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48575.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48790.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48791.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48801.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] AAN86226.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48840.1 MSP1 major blood-stage surface antigen Pv200 [Plasmodium vivax Sal-1] XP_001614842.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48653.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48552.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48793.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] AAN86235.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48836.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48574.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48811.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48826.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48822.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48557.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] AAN86209.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] AAN86220.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48590.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48794.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] AAN86212.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48555.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48579.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48547.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48816.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48602.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48591.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] AAN86223.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] AAN86227.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48824.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] AAN86213.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48796.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48804.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] AAN86228.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48542.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48563.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48831.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48819.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] AAN86248.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] AAN86236.1 MSP1 merozoite surface protein-1 [Plasmodium vivax] ADF48784.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] AAN86215.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] AAN86214.1 MSP1 merozoite surface protein 1 [Plasmodium vivax] ABB46193.1 MSP1 merozoite surface protein 1 [Plasmodium Malariae] ACZ51237.1 MSP1 merozoite surface protein 1 [Plasmodium ovale] ACZ51238.1 MSP1 merozoite surface protein-1 [Plasmodium ovale] AGI42775.1 MSP1 merozoite surface protein 1 [Plasmodium ovale] ACZ51239.1 MSP1 merozoite surface protein-1 [Plasmodium ovale] AGI42781.1 MSP1 merozoite surface protein-1 [Plasmodium ovale] AGI42772.1 MSP1 merozoite surface protein-1 [Plasmodium ovale] AGI42773.1 MSP1 merozoite surface protein-1 [Plasmodium ovale] AGI42777.1 MSP2 merozoite surface protein 1 [Plasmodium vivax] AAN86237.1 MSP3 merozoite surface protein-1 [Plasmodium vivax] ADF48815.1 RIFIN RIFIN [Plasmodium falciparum] ABB72339.1 RIFIN hypothetical protein PFMAFIP_05646 [Plasmodium falciparum ETW46404.1 MaliPS096_E11] RIFIN hypothetical protein PFAG_05918 [Plasmodium falciparum Santa Lucia] EUT78013.1 RIFIN hypothetical protein PFNF135_05454 [Plasmodium falciparum NF135/5.C10] ETW40226.1 RIFIN rifin [Plasmodium falciparum IGH-CR14] KNG78559.1 RIFIN hypothetical protein PFFVO_05462 [Plasmodium falciparum Vietnam Oak- ETW15682.1 Knoll (FVO)] RIFIN rifin [Plasmodium falciparum 3D7] XP_001350919.1 RIFIN RIFIN [Plasmodium falciparum] ABB72268.1 RIFIN rifin [Plasmodium falciparum IGH-CR14] KNG75134.1 RIFIN rifin [Plasmodium reichenowi] XP_012760698.1 SSP2 sporozoite surface protein 2 [Plasmodium vivax] AAC97485.1 SSP2 sporozoite surface protein 2 [Plasmodium vivax] AAC97484.1 SSP2 sporozoite surface protein 2 [Plasmodium vivax Sal-1] XP_001614147.1 SSP2 sporozoite surface protein 2 [Plasmodium vivax India VII] KMZ79303.1 SSP2 sporozoite surface protein 2 [Plasmodium vivax Mauritania I] KMZ91322.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] BAO53712.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] BAO53686.1 TRAP hypothetical protein PFNF135_04795 [Plasmodium falciparum NF135/5.C10] ETW40491.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] BAO53689.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] BAO53710.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] BAO53701.1 TRAP thrombospondin-related protein [Plasmodium falciparum] BAA31169.1 TRAP thrombospondin-related protein [Plasmodium falciparum] BAA31194.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] BAO53705.1 TRAP thrombospondin-related protein [Plasmodium falciparum] BAA31175.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] BAO53708.1 TRAP Thrombospondin-related anonymous protein, TRAP [Plasmodium falciparum XP_001350088.1 3D7] TRAP thrombospondin-related protein [Plasmodium falciparum] BAA31182.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] BAO53682.1 TRAP thrombospondin related anonymous protein [Plasmodium falciparum] AAA29775.1 TRAP thrombospondin-related protein [Plasmodium falciparum] BAA31176.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] BAO53709.1 TRAP thrombospondin-related protein [Plasmodium falciparum] BAA31183.1 TRAP thrombospondin related anonymous protein [Plasmodium falciparum] AAA29773.1 TRAP thrombospondin-related protein [Plasmodium falciparum] BAA31168.1 TRAP sporozoite surface protein 2 [Plasmodium falciparum] AAG12328.1 TRAP thrombospondin related anonymous protein [Plasmodium falciparum] AAA29776.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAQ11895.1 TRAP thrombospondin related anonymous protein [Plasmodium falciparum] AAA29774.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] BA053700.1 TRAP thrombospondin-related protein [Plasmodium falciparum] BAA31174.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] BAO53711.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] BAO53684.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] BAO53683.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] BAO53698.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] BAO53687.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] BAO53707.1 TRAP thrombospondin related anonymous protein [Plasmodium falciparum] AAA29778.1 TRAP thrombospondin-related protein [Plasmodium falciparum] BAA31186.1 TRAP thrombospondin-related protein [Plasmodium falciparum] BAA31173.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAQ11894.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] BAO53691.1 TRAP thrombospondin-related protein [Plasmodium falciparum] BAA31191.1 TRAP thrombospondin related anonymous protein [Plasmodium falciparum] AAA29772.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] BAO53695.1 TRAP thrombospondin-related protein [Plasmodium falciparum] BAA31190.1 TRAP thrombospondin-related protein [Plasmodium falciparum] BAA31188.1 TRAP thrombospondin-related protein [Plasmodium falciparum] BAA31192.1 TRAP thrombospondin-related protein [Plasmodium falciparum] BAA31180.1 TRAP sporozoite surface protein 2 [Plasmodium falciparum IGH-CR14] KNG77170.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAQ11892.1 TRAP sporozoite surface protein 2 [Plasmodium falciparum RAJ116] KNC36192.1 TRAP thrombospondin-related protein [Plasmodium falciparum] BAA31171.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] BAO53704.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAQ11891.1 TRAP thrombospondin related anonymous protein [Plasmodium falciparum] AAA29771.1 TRAP thrombospondin-related protein [Plasmodium falciparum] BAA31177.1 TRAP thrombospondin-related protein [Plasmodium falciparum] BAA31181.1 TRAP thrombospondin-related protein [Plasmodium falciparum] BAA31170.1 TRAP thrombospondin-related protein [Plasmodium falciparum] BAA31189.1 TRAP thrombospondin-related protein [Plasmodium falciparum] BAA31193.1 TRAP hypothetical protein PFUGPA_02262 [Plasmodium falciparum Palo ETW55500.1 Alto/Uganda] TRAP thrombospondin-related protein [Plasmodium falciparum] BAA31172.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAW78134.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAW78169.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAW78171.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAW78149.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAW78151.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAW78161.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAW78144.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAW78137.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAW78153.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAW78163.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAW78135.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAW78166.1 TRAP thrombospondin-related protein [Plasmodium falciparum] BAA31187.1 TRAP thrombospondin-related protein [Plasmodium falciparum] BAA31178.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] BAO53702.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAW78172.1 TRAP thrombospondin related anonymous protein [Plasmodium falciparum] AAA29777.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAW78152.1 TRAP thrombospondin related anonymous protein [Plasmodium falciparum] AAA29770.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAW78168.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAW78159.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAW78155.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAW78148.1 TRAP thrombospondin-related protein [Plasmodium falciparum] BAA31167.1 TRAP hypothetical protein PFFCH_03096 [Plasmodium falciparum FCH/4] ETW29444.1 TRAP Thrombospondin related anonymous protein [Plasmodium cynomolgi] CAA63617.1 TRAP hypothetical protein C923_04716 [Plasmodium falciparum UGT5.1] EWC74605.1 TRAP RecName: Full = Thrombospondin-related anonymous protein; Flags: P16893.1 Precursor TRAP thrombospondin related protein TRAP [Plasmodium falciparum] 1411304A TRAP thrombospondin related protein [Plasmodium falciparum] 1708291A TRAP hypothetical protein PFMAFIP_04474 [Plasmodium falciparum ETW47461.1 MaliPS096_E11] TRAP hypothetical protein PFBG_04660 [Plasmodium falciparum 7G8] EUR65920.1 TRAP thrombospondin related adhesive protein [Plasmodium falciparum] AAC18657.1 TRAP hypothetical protein PFTANZ_06525 [Plasmodium falciparum Tanzania ETW32755.1 (2000708)] TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAW78160.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAW78131.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAW78130.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAW78143.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAW78164.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAW78165.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAW78167.1 TRAP thrombospondin-related adhesive protein [Plasmodium falciparum] AAW78146.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97094.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97016.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97071.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97015.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97087.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97084.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97050.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97019.1 TRAP sporozoite surface protein 2 [Plasmodium vivax Mauritania I] KMZ91322.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97011.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97045.1 TRAP sporozoite surface protein 2 [Plasmodium vivax India VII] KMZ79303.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97046.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97013.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97061.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97065.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97037.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97029.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97110.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97026.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97023.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97058.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97006.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97075.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97027.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97066.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97063.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97040.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97004.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97003.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97034.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97005.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97070.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97069.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97012.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97020.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97053.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97041.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97014.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97008.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97052.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AIU97035.1 TRAP thrombospondin-related anonymous protein [Plasmodium vivax] AAC47463.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57608.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57598.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57607.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57595.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57588.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57580.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57567.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57599.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57597.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57592.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57638.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57606.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57585.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57581.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57629.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57593.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57605.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57578.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57631.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57570.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57628.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57611.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57573.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57579.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57634.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57576.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57591.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57584.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57577.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57600.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57620.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57603.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57621.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57604.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57583.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57582.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57632.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57617.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57590.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57575.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57630.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57623.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57612.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57610.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57601.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57624.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57637.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57619.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57639.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57636.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AAK57618.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AEC32940.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AEC32938.1 TRAP thrombospondin-related adhesive protein [Plasmodium vivax] AEC32936.1

TABLE 4 JEV Nucleic Acid Sequences Antigen Nucleic Acid Sequence SEQ ID NO: Japanese ATGCTTGGCAGCAATAGTGGTCAACGTGTGGTGTTCACTATCCTC 18 encephalitis CTGCTGTTGGTCGCTCCGGCTTACAGTTTTAACTGTCTGGGAATG virus, Thailand, GGGAATCGAGATTTCATAGAAGGAGCCAGTGGAGCCACTTGGGTG strain: 4790-85, GATCTGGTATTAGAAGGAGACAGTTGTTTGACAATCATGGCAAAC envelope GACAAACCAACACTAGATGTCCGCATGATCAACATTGAAGCCAGC glycoprotein CAACTCGCTGAAGTTAGGAGCTACTGCTATCACGCTTCAGTCACT E or E GACATTTCAACGGTGGCTCGATGCCCCACGACTGGAGAAGCTCAC AACGAGAAACGTGCTGACAGCAGCTACGTGTGCAAGCAAGGCTTT ACTGACCGCGGATGGGGAAATGGATGTGGACTTTTCGGGAAAGGA AGCATTGACACTTGCGCAAAGTTTACTTGTACTAATAAGGCCATT GGAAGAACAATCCAACCAGAGAACATCAAGTATGAAGTTGGTGTA TTCGTGCACGGAACCACCACCTCGGAAAACCATGGGAATTACTCA GCGCAGGTTGGAGCGTCTCAAGCAGCAAAGTTTACTGTAACTCCA AACGCTCCCTCAATAACCCTCAAGCTTGGTGATTATGGAGAAGTT ACACTGGATTGTGAACCAAGGAGTGGACTGAACACTGAAGCGTTC TATGTCATGACTGTGGGTTCGAAGTCATTCTTAGTCCATAGGGAA TGGTTCCATGACCTTTCTCTTCCCTGGACATCCCCCTCAAGCACG GCATGGAGGAACAGAGAACTCCTTATGGAATTTGAAGAGGCACAT GCCACAAAACAATCTGTCGTAGCCCTTGGGTCACAGGAGGGAGGC CTCCATCAAGCGTTAGCAGGAGCCATCGTGGTGGAGTACTCAAGC TCAGTGAAGTTGACATCAGGTCACCTGAAATGCAGGCTGAAAATG GACAAACTGGCTCTGAAGGGCACGACTTATGGCATGTGCACAGAA AAATTCTCGTTCGCGAAAAACCCAGCGGACACAGGCCATGGAACA GTTGTCATTGAGCTCACATACTCTGGAAGCGATGGTCCCTGCAAA ATTCCGATTGTTTCAGTCGCGAGTTTAAACGACATGACCCCTGTG GGAAGGCTGGTAACAGTAAACCCCTTCGTCGCGACATCCAGCTCC AACTCGAAGGTGCTGGTTGAGATGGAACCTCCTTTCGGAGACTCT TATATTGTGGTTGGAAGAGGGGACAAGCAGATTAACCATCACTGG CACAAAGCTGGAAGCACGTTGGGCAAAGCCTTCTCAACAACTTTG AAAGGAGCTCAAAGACTGGCAGCGCTAGGGGACACAGCCTGGGAC TTTGGTTCCATTGGGGGGGTACTCAACTCCATAGGAAAAGCTGTT CACCAAGTATTTGGCGGTGCATTTAGAACGCTTTTTGGGGGAATG TCTTGGATCACGCAAGGGCTAATGGGGGCCCTACTTCTCTGGATG GGTGTCAACGCACGAGATCGGTCAATCGCCTTAGCTTTTTTGGCC ACGGGAGGTGTGCTCGTCTTTTTAGCGACCAACGTGCATGCT Japanese ATGTGGCTTGTCAGCTTGGCAATCGTAACAGCTTGTGCCGGAGCT 19 encephalitis ATGAAGCTATCAAACTTTCAAGGAAAGCTTCTGATGACCATCAAC virus, Thailand, AACACGGACATTGCGGACGTCATCGTGATCCCTACCTCAAAAGGC strain: 4790-85, GAAAACAGATGTTGGGTCCGAGCGATCGACGTTGGTTACATGTGT envelope GAAGACACCATTACGTACGAATGTCCGAAGCTAGCAGTGGGCAAC glycoprotein M GACCCAGAAGACGTGGATTGCTGGTGCGACAATCAAGAAGTCTTC and envelope GTGCAGTATGGTCGCTGCACACGGACCAGGCATTCCAAACGAAGC glycoprotein E AGAAGATCCGTTTCGGTCCAAACGCATGGTGAAAGCTCACTAGTG polyprotein AACAAAAAAGAGGCTTGGCTGGATTCAACGAAGGCCACGCGATAC Or PrM-E CTCATGAAAACGGAGAATTGGATCATAAGGAACCCTGGTTATGCT TTCCTGGCGGCGGCACTTGGTTGGATGCTTGGCAGCAATAGTGGT CAACGTGTGGTGTTCACTATCCTCCTGCTGTTGGTCGCTCCGGCT TACAGTTTTAACTGTCTGGGAATGGGGAATCGAGATTTCATAGAA GGAGCCAGTGGAGCCACTTGGGTGGATCTGGTATTAGAAGGAGAC AGTTGTTTGACAATCATGGCAAACGACAAACCAACACTAGATGTC CGCATGATCAACATTGAAGCCAGCCAACTCGCTGAAGTTAGGAGC TACTGCTATCACGCTTCAGTCACTGACATTTCAACGGTGGCTCGA TGCCCCACGACTGGAGAAGCTCACAACGAGAAACGTGCTGACAGC AGCTACGTGTGCAAGCAAGGCTTTACTGACCGCGGATGGGGAAAT GGATGTGGACTTTTCGGGAAAGGAAGCATTGACACTTGCGCAAAG TTTACTTGTACTAATAAGGCCATTGGAAGAACAATCCAACCAGAG AACATCAAGTATGAAGTTGGTGTATTCGTGCACGGAACCACCACC TCGGAAAACCATGGGAATTACTCAGCGCAGGTTGGAGCGTCTCAA GCAGCAAAGTTTACTGTAACTCCAAACGCTCCCTCAATAACCCTC AAGCTTGGTGATTATGGAGAAGTTACACTGGATTGTGAACCAAGG AGTGGACTGAACACTGAAGCGTTCTATGTCATGACTGTGGGTTCG AAGTCATTCTTAGTCCATAGGGAATGGTTCCATGACCTTTCTCTT CCCTGGACATCCCCCTCAAGCACGGCATGGAGGAACAGAGAACTC CTTATGGAATTTGAAGAGGCACATGCCACAAAACAATCTGTCGTA GCCCTTGGGTCACAGGAGGGAGGCCTCCATCAAGCGTTAGCAGGA GCCATCGTGGTGGAGTACTCAAGCTCAGTGAAGTTGACATCAGGT CACCTGAAATGCAGGCTGAAAATGGACAAACTGGCTCTGAAGGGC ACGACTTATGGCATGTGCACAGAAAAATTCTCGTTCGCGAAAAAC CCAGCGGACACAGGCCATGGAACAGTTGTCATTGAGCTCACATAC TCTGGAAGCGATGGTCCCTGCAAAATTCCGATTGTTTCAGTCGCG AGTTTAAACGACATGACCCCTGTGGGAAGGCTGGTAACAGTAAAC CCCTTCGTCGCGACATCCAGCTCCAACTCGAAGGTGCTGGTTGAG ATGGAACCTCCTTTCGGAGACTCTTATATTGTGGTTGGAAGAGGG GACAAGCAGATTAACCATCACTGGCACAAAGCTGGAAGCACGTTG GGCAAAGCCTTCTCAACAACTTTGAAAGGAGCTCAAAGACTGGCA GCGCTAGGGGACACAGCCTGGGACTTTGGTTCCATTGGGGGGGTA CTCAACTCCATAGGAAAAGCTGTTCACCAAGTATTTGGCGGTGCA TTTAGAACGCTTTTTGGGGGAATGTCTTGGATCACGCAAGGGCTA ATGGGGGCCCTACTTCTCTGGATGGGTGTCAACGCACGAGATCGG TCAATCGCCTTAGCTTTTTTGGCCACGGGAGGTGTGCTCGTCTTT TTAGCGACCAACGTGCATGCT JEV mRNA Sequences Japanese AUGCUUGGCAGCAAUAGUGGUCAACGUGUGGUGUUCACUAUCCUC 20 encephalitis CUGCUGUUGGUCGCUCCGGCUUACAGUUUUAACUGUCUGGGAAUG virus, Thailand, GGGAAUCGAGAUUUCAUAGAAGGAGCCAGUGGAGCCACUUGGGUG strain: 4790-85, GAUCUGGUAUUAGAAGGAGACAGUUGUUUGACAAUCAUGGCAAAC envelope GACAAACCAACACUAGAUGUCCGCAUGAUCAACAUUGAAGCCAGC glycoprotein CAACUCGCUGAAGUUAGGAGCUACUGCUAUCACGCUUCAGUCACU E or E GACAUUUCAACGGUGGCUCGAUGCCCCACGACUGGAGAAGCUCAC AACGAGAAACGUGCUGACAGCAGCUACGUGUGCAAGCAAGGCUUU ACUGACCGCGGAUGGGGAAAUGGAUGUGGACUUUUCGGGAAAGGA AGCAUUGACACUUGCGCAAAGUUUACUUGUACUAAUAAGGCCAUU GGAAGAACAAUCCAACCAGAGAACAUCAAGUAUGAAGUUGGUGUA UUCGUGCACGGAACCACCACCUCGGAAAACCAUGGGAAUUACUCA GCGCAGGUUGGAGCGUCUCAAGCAGCAAAGUUUACUGUAACUCCA AACGCUCCCUCAAUAACCCUCAAGCUUGGUGAUUAUGGAGAAGUU ACACUGGAUUGUGAACCAAGGAGUGGACUGAACACUGAAGCGUUC UAUGUCAUGACUGUGGGUUCGAAGUCAUUCUUAGUCCAUAGGGAA UGGUUCCAUGACCUUUCUCUUCCCUGGACAUCCCCCUCAAGCACG GCAUGGAGGAACAGAGAACUCCUUAUGGAAUUUGAAGAGGCACAU GCCACAAAACAAUCUGUCGUAGCCCUUGGGUCACAGGAGGGAGGC CUCCAUCAAGCGUUAGCAGGAGCCAUCGUGGUGGAGUACUCAAGC UCAGUGAAGUUGACAUCAGGUCACCUGAAAUGCAGGCUGAAAAUG GACAAACUGGCUCUGAAGGGCACGACUUAUGGCAUGUGCACAGAA AAAUUCUCGUUCGCGAAAAACCCAGCGGACACAGGCCAUGGAACA GUUGUCAUUGAGCUCACAUACUCUGGAAGCGAUGGUCCCUGCAAA AUUCCGAUUGUUUCAGUCGCGAGUUUAAACGACAUGACCCCUGUG GGAAGGCUGGUAACAGUAAACCCCUUCGUCGCGACAUCCAGCUCC AACUCGAAGGUGCUGGUUGAGAUGGAACCUCCUUUCGGAGACUCU UAUAUUGUGGUUGGAAGAGGGGACAAGCAGAUUAACCAUCACUGG CACAAAGCUGGAAGCACGUUGGGCAAAGCCUUCUCAACAACUUUG AAAGGAGCUCAAAGACUGGCAGCGCUAGGGGACACAGCCUGGGAC UUUGGUUCCAUUGGGGGGGUACUCAACUCCAUAGGAAAAGCUGUU CACCAAGUAUUUGGCGGUGCAUUUAGAACGCUUUUUGGGGGAAUG UCUUGGAUCACGCAAGGGCUAAUGGGGGCCCUACUUCUCUGGAUG GGUGUCAACGCACGAGAUCGGUCAAUCGCCUUAGCUUUUUUGGCC ACGGGAGGUGUGCUCGUCUUUUUAGCGACCAACGUGCAUGCU Japanese AUGUGGCUUGUCAGCUUGGCAAUCGUAACAGCUUGUGCCGGAGCU 21 encephalitis AUGAAGCUAUCAAACUUUCAAGGAAAGCUUCUGAUGACCAUCAAC virus, Thailand, AACACGGACAUUGCGGACGUCAUCGUGAUCCCUACCUCAAAAGGC strain: 4790-85, GAAAACAGAUGUUGGGUCCGAGCGAUCGACGUUGGUUACAUGUGU envelope GAAGACACCAUUACGUACGAAUGUCCGAAGCUAGCAGUGGGCAAC glycoprotein M GACCCAGAAGACGUGGAUUGCUGGUGCGACAAUCAAGAAGUCUUC and envelope GUGCAGUAUGGUCGCUGCACACGGACCAGGCAUUCCAAACGAAGC glycoprotein E AGAAGAUCCGUUUCGGUCCAAACGCAUGGUGAAAGCUCACUAGUG polyprotein AACAAAAAAGAGGCUUGGCUGGAUUCAACGAAGGCCACGCGAUAC Or PrM-E CUCAUGAAAACGGAGAAUUGGAUCAUAAGGAACCCUGGUUAUGCU UUCCUGGCGGCGGCACUUGGUUGGAUGCUUGGCAGCAAUAGUGGU CAACGUGUGGUGUUCACUAUCCUCCUGCUGUUGGUCGCUCCGGCU UACAGUUUUAACUGUCUGGGAAUGGGGAAUCGAGAUUUCAUAGAA GGAGCCAGUGGAGCCACUUGGGUGGAUCUGGUAUUAGAAGGAGAC AGUUGUUUGACAAUCAUGGCAAACGACAAACCAACACUAGAUGUC CGCAUGAUCAACAUUGAAGCCAGCCAACUCGCUGAAGUUAGGAGC UACUGCUAUCACGCUUCAGUCACUGACAUUUCAACGGUGGCUCGA UGCCCCACGACUGGAGAAGCUCACAACGAGAAACGUGCUGACAGC AGCUACGUGUGCAAGCAAGGCUUUACUGACCGCGGAUGGGGAAAU GGAUGUGGACUUUUCGGGAAAGGAAGCAUUGACACUUGCGCAAAG UUUACUUGUACUAAUAAGGCCAUUGGAAGAACAAUCCAACCAGAG AACAUCAAGUAUGAAGUUGGUGUAUUCGUGCACGGAACCACCACC UCGGAAAACCAUGGGAAUUACUCAGCGCAGGUUGGAGCGUCUCAA GCAGCAAAGUUUACUGUAACUCCAAACGCUCCCUCAAUAACCCUC AAGCUUGGUGAUUAUGGAGAAGUUACACUGGAUUGUGAACCAAGG AGUGGACUGAACACUGAAGCGUUCUAUGUCAUGACUGUGGGUUCG AAGUCAUUCUUAGUCCAUAGGGAAUGGUUCCAUGACCUUUCUCUU CCCUGGACAUCCCCCUCAAGCACGGCAUGGAGGAACAGAGAACUC CUUAUGGAAUUUGAAGAGGCACAUGCCACAAAACAAUCUGUCGUA GCCCUUGGGUCACAGGAGGGAGGCCUCCAUCAAGCGUUAGCAGGA GCCAUCGUGGUGGAGUACUCAAGCUCAGUGAAGUUGACAUCAGGU CACCUGAAAUGCAGGCUGAAAAUGGACAAACUGGCUCUGAAGGGC ACGACUUAUGGCAUGUGCACAGAAAAAUUCUCGUUCGCGAAAAAC CCAGCGGACACAGGCCAUGGAACAGUUGUCAUUGAGCUCACAUAC UCUGGAAGCGAUGGUCCCUGCAAAAUUCCGAUUGUUUCAGUCGCG AGUUUAAACGACAUGACCCCUGUGGGAAGGCUGGUAACAGUAAAC CCCUUCGUCGCGACAUCCAGCUCCAACUCGAAGGUGCUGGUUGAG AUGGAACCUCCUUUCGGAGACUCUUAUAUUGUGGUUGGAAGAGGG GACAAGCAGAUUAACCAUCACUGGCACAAAGCUGGAAGCACGUUG GGCAAAGCCUUCUCAACAACUUUGAAAGGAGCUCAAAGACUGGCA GCGCUAGGGGACACAGCCUGGGACUUUGGUUCCAUUGGGGGGGUA CUCAACUCCAUAGGAAAAGCUGUUCACCAAGUAUUUGGCGGUGCA UUUAGAACGCUUUUUGGGGGAAUGUCUUGGAUCACGCAAGGGCUA AUGGGGGCCCUACUUCUCUGGAUGGGUGUCAACGCACGAGAUCGG UCAAUCGCCUUAGCUUUUUUGGCCACGGGAGGUGUGCUCGUCUUU UUAGCGACCAACGUGCAUGCU

TABLE 5 JEV Amino Acid Sequences Accession Amino Acid Sequence SEQ ID NO: gi|307826672|gb| MLGSNSGQRVVFTILLLLVAPAYSFNCLGMGNRDFIEGASGATWV 22 ADN94470.1| DLVLEGDSCLTIMANDKPTLDVRMINIEASQLAEVRSYCYHASIN polyprotein VTDISTVARCPTTGEAHNEKRADSSYVCKQGFTDRGWGNGCGLFG [Japanese KGSIDTCAKFTCTNKAIGRTIQPENIKYEVGVFVHGTTTSENHGN encephalitis YSAQVGASQAAKFTVTPNAPSITLKLGDYGEVTLDCEPRSGLNTE virus] AFYVMTVGSKSFLVHREWFHDLSLPWTSPSSTAWRNRELLMEFEE Envelope AHATKQSINVVALGSQEGGLHQALAGAIVVEYSSSINVKLTSGHL glycoprotein KCRLKMDKLALKGTTYGMCTEKFSFAKNPADTGHGTVVIELTYSG E or E SDGPCKIPIVSINVASLNDMTPVGRLVTVNPFVATSSSNSKVLVE MEPPFGDSYIVVGRGDKQINHHWHKAGSTLGKAFSTTLKGAQRLA ALGDTAWDFGSIGGVLNSIGKAVHQVFGGAFRTLFGGMSWITQGL MGALLLWMGVNARDRSIALAFLATGGVLVFLATNVHA gi|307826672|gb| MWLVSLAIVTACAGAMKLSNFQGKLLMTINNTDIADVIVIPTSKG 23 ADN94470.1| ENRCWVRAIDVGYMCEDTITYECPKLAVGNDPEDVDCWCDNQEVF polyprotein VQYGRCTRTRHSKRSRRSINVSINVQTHGESSLVNKKEAWLDSTK envelope ATRYLMKTENWIIRNPGYAFLAAALGWMLGSNSGQRVVFTILLLL glycoprotein M VAPAYSFNCLGMGNRDFIEGASGATWVDLVLEGDSCLTIMANDKP and envelope TLDVRMINIEASQLAEVRSYCYHASINVTDISTVARCPTTGEAHN glycoprotein E EKRADSSYVCKQGFTDRGWGNGCGLFGKGSIDTCAKFTCTNKAIG polyprotein RTIQPENIKYEVGVFVHGTTTSENHGNYSAQVGASQAAKFTVTPN OR JEV_PrM-E APSITLKLGDYGEVTLDCEPRSGLNTEAFYVMTVGSKSFLVHREW FHDLSLPWTSPSSTAWRNRELLMEFEEAHATKQSINVVALGSQEG GLHQALAGAIVVEYSSSINVKLTSGHLKCRLKMDKLALKGTTYGM CTEKFSFAKNPADTGHGTVVIELTYSGSDGPCKIPIVSINVASLN DMTPVGRLVTVNPFVATSSSNSKVLVEMEPPFGDSYIVVGRGDKQ INHHWHKAGSTLGKAFSTTLKGAQRLAALGDTAWDFGSIGGVLNS IGKAVHQVFGGAFRTLFGGMSWITQGLMGALLLWMGVNARDRSIA LAFLATGGVLVFLATNVHA >gi|126508804| MTKKPGGPGKNRAINMLKRGLPRVFPLVGVKRVVMSLLDGRGPVR 24 gb|ABO15575.1 FVLALITFFKFTALAPTKALLGRWRAVEKSINVAMKHLTSFKREL 1 polyprotein GTLIDAVNKRGKKQNKRGGNESSIMWLVSLAIVTACAGAMKLSNF QGKLLMTINNTDIADVIVIPTSKGENRCWVRAIDVGYMCEDTITY ECPKLAVGNDPEDVDCWCDNQEVFVQYGRCTRTRHSKRSRRSINV SINVQTHGESSLVNKKEAWLDSTKATRYLMKTENWIIRNPGYAFL AAALGWMLGSNSGQRVVFTILLLLVAPAYSFNCLGMGNRDFIEGA SGATWVDLVLEGDSCLTIMANDKPTLDVRMINIEASQLAEVRSYC YHASINVTDISTVARCPTTGEAHNEKRADSSYVCKQGFTDRGWGN GCGLFGKGSIDTCAKFTCNNKAIGRTIQPENIKYEVGVFVHGTTT SENHGNYSAQVGASQAAKFTVTPNAPSITLKLGDYGEVTLDCEPR SGLNTEAFYVMTVGSKSFLVHREWFHDLSLPWTSPSSTAWRNREL LMEFEEAHATKQSINVVALGSQEGGLHQALAGAIVVEYSSSINVK LTSGHLKCRLKMDKLALKGTTYGMCTEKFSFAKNPADTGHGTVVI ELTYSGSDGPCKIPIVSINVASLNDMTPVGRLVTVNPFVATSSSN SKVLVEMEPPFGDSYIVVGRGDKQINHHWHKAGSTLGKAFSTTLK GAQRLAALGDTAWDFGSIGGVLNSIGKAVHQVFGGAFRTLFGGMS WITQGLMGALLLWMGVNARDRSIALAFLATGGVLVFLATNVHADT GCAIDITRKEMRCGS >bi|824555713| MTKKPGGPGKNRAINMLKRGLPRVFPLVGVKRVVMSLLDGRGPVR 25 dbj|BAR88119.1| FVLALITFFKFTALAPTKALLGRWRAVEKSINVAMKHLTSFKREL polyprotein GTLIDAVNKRGKKQNKRGGNESSIMWLASLAIVTACAGAMKLSNF QGKLLMTINNTDIADVIVIPTSKGENRCWVRAIDVGYMCEDTITY ECPKLAVGNDPEDVDCWCDNQEVYVQYGRCTRTRHSKRSRRSINV SINVQTHGESSLVNKKEAWLDSTKATRYLMKTENWIIRNPGYAFL AAALGWMLGSNSGQRVVFTILLLLVAPAYSFNCLGMGNRDFIEGA SGATWVDLVLEGDSCLTIMANDKPTLDVRMINIEASQLAEVRSYC YHASINVTDISTVARCPTTGEAHNEKRADSSYVCKQGFTDRGWGN GCGLFGKGSIDTCAKFSCTNKAIGRMIQPENIKYEVGIFVHGTTT SENHGNYSAQVGASQAAKFTVTPNAPSITLKLGDYGEVTLDCEPR SGLNTEAFYVMTVGSKSFLVHREWFHDLSLPWTSPSSTAWRNREL LMEFEEAHATKQSINVVALGSQEGGLHQALAGAIVVEYSSSINVK LTSGHLKCRLKMDKLALKGTTYGMCTEKFSFAKNPADTGHGTVVI ELTYSGSDGPCKIPIVSINVASLNDMTPVGRLVTVNPFVATSSSN SKVLVEMEPPFGDSYIVVGRGDKQINHHWHKAGSTLGKAFSTTLK GAQRLAALGDTAWDFGSIGGVFNSIGKAVHQVFGGAFRTLFGGMS WITQGLMGALLLWMGVNARDRSIALAFLATGGVLVFLATNVHADT GCAIDITRKEMRCGSGIFVHNDVEAWVDRYKYLPETPRSLAKIVH KAHQEGVCGVRSINVTRLEHQMWESINVRDELNVLLKENAVDLSI NVVVNKPVGRYRSAPKRLSMTQEKFEMGWKAWGKSILFAPELANS TFVVDGPETKECPDERRAWNSMQIEDFGFGITSTRVWLKIREENT DECDGAIIGTAVKGHVAVHSDLSYWIESRLNDTWKLERAVFGEVK SCTWPETHTLWGDGVEESELIIPHTIAGPRSKHNRREGYKTQNQG PWDENGIVLDFDYCPGTKVTITEDCGKRGPSIRTTTDSGKLITDW CCRSCSLPPLRFRTENGCWYGMEIRPVRHDETTLVRSQVDAFNGE MIDPFQLGLLVMFLATQEVLRKRWTARLTIPAVLGALLVLMLGGI TYTDMARYVVLVAAAFAEANSGGDVLHLALIAVFKIQPSFLVMNM LSARWTNQENVVLVLGAAFFQLASINVDLQIGVHGILNAAAIAWM IVRAITFPTTSTVAMPVLALLTPGMRALYLDTYRIILLVIGICSL LQERRKTMAKKKGAVLLGLALTSTGWFSPTTIAAGLMVCNPNKKR GWPATEFLSAVGLMFAIVGGLAELDIESMSIPFMLAGLMAVSYVV SGKATDMWLDRAADISWEMEAAITGSSRRLDVKLDDDGDFHLIDD PGVPWKVWLLRMSCIGLAALTPWAIVPAAFGYWLTLKTTKRGGVF WDTPSPKPCLKGDTTTGVYRIMARGILGTYQAGVGVMYENVFHTL WHTTRGAAIMSGEGKLTPYWGSINVKEDRISYGGPWRFDRKWNGT DDVQVIVVEPGKPAVNIQTKPGVFRTPFGEIGAVSLDYPRGTSGS PILDSNGDIIGLYGNGVELGDGSYVSAIVQGDRQEEPVPDAYTPG MLKKRQMTVLDLHPGSGKTRKILPQIIKDAIQQRLRTAVLAPTRV VAAEMAEALKGLPVRYQTSAVQREHQGNEIVDVMCHATLTHRLMS PNRVPNYNLFVMDEAHFTDPASIAARGYIATKVELGEAAAIFMTA TPPGTTDPFPDSNAPIHDLQDEIPDRAWSSGYEWITDYAGKTVWF VASINVKMGNEIAMCLQRAGKKVIQLNRKSYDTEYPKCKNGDWDF VITTDISEMGANFGASRVIDCRKSINVKPTILEEGEGRVILGNPS PITSASAAQRRGRVGRNPNQVGDEYHYGGATSEDDSNLAHWTEAK IMLDNIHMPNGLVAQLYGPEREKAFTMDGEYRLRGEEKKNFLELL RTADLPVWLAYKVASNGIQYTDRKWCFDGPRTNAILEDNTEVEIV TRMGERKILKPRWLDARVYADHQALKWFKDFAAGKRSAVSFIEVL GRMPEHFMGKTREALDTMYLVATAEKGGKAHRMALEELPDALETI TLIVAITVMTGGFFLLMMQRKGIGKMGLGALVLTLATFFLWAAEV PGTKIAGTLLVALLLMVVLIPEPEKQRSQTDNQLAVFLICVLTVV GVVAANEYGMLEKTKADLKSMFGGRTQASGLTGLPSMALDLRPAT AWALYGGSTVVLTPLLKHLITSEYVTTSLASISSQAGSLFVLPRG VPFTDLDLTVGLVFLGCWGQITLTTFLTAMVLVTLHYGYMLPGWQ AEALRAAQRRTAAGIMKNAVVDGMVATDVPELERTTPLMQKKVGQ VLLIGVSINVAAFLVNPNVTTVREAGVLVTAATLTLWDNGASAVW NSTTATGLCHVMRGSYLAGGSIAWTLIKNADKPSLKRGRPGGRTL GEQWKEKLNAMSRDEFFKYRREAIIEVDRTEARRARRENNIVGGH PVSRGSAKLRWLVEKGFVSPIGKVIDLGCGRGGWSYYAATLKKVQ EVKGYTKGGAGHEEPMLMQSYGWNLVSLKSGVDVFYKPSEPSDTL FCDIGESSPSPEVEEQRTLRVLEMTSDWLHRGPREFCIKVLCPYM PKVIEKMEVLQRRFGGGLVRLPLSRNSNHEMYWVSGAAGNVVHAV NMTSQVLLGRMDRTVWRGPKYEEDVNLGSGTRAVGKGEVHSNQEK IRKRIQKLKEEFATTWHKDPEHPYRTWTYHGSYEVKATGSASSLV NGVVKLMSKPWDAIANVTTMAMTDTTPFGQQRVFKEKVDTKAPEP PAGVKEVLNETTNWLWAHLSREKRPRLCTKEEFIKKVNSNAALGA VFAEQNQWSTAREAVGDPLFWEMVNEERENHLRGECHTCIYNMMG KREKKPGEFGKAKGSRAIWFMWLGARYLEFEALGFLNEDHWLSRE NSGGGVEGSGVQKLGYILRDIAGKQGGKMYADDTAGWDTRITRTD LENEAKVLELLDGEHRMLARAIIELTYRHKVVKVMRPAAGGKTVM DVISREDQRGSGQVVTYALNTFTNIAVQLVRLMEAEGVIGPQHLE QLPRKNKIAVRTWLFENGEERVTRMAISGDDCVVKPLDDRFATAL HFLNAMSKVRKDIQEWKPSHGWHDWQQVPFCSNHFQEIVMKDGRS IVVPCRGQDELIGRARISPGAGWNVKDTACLAKAYAQMWLLLYFH RRDLRLMANAICSAVPVDWVPTGRTSWSIHSKGEWMTTEDMLQVW NRVWIEENEWMMDKTPITSWTDVPYVGKREDIWCGSLIGTRSRAT WAENVYAAINQVRAIIGKENYVDYMTSLRRYEDVLIQEDRVI >gi|824555711| MTKKPGGPGKNRAINMLKRGLPRVFPLVGVKRVVMSLLDGRGPVR 26 dbj|BAR88118.1| FVLALITFFKFTALAPTKALLGRWRAVEKSINVAMKHLTSFKREL polyprotein GTLIDAVNKRGKKQNKRGGNESSIVWLASLAIVTACAGAMKLSNF QGKLLMTINNTDIADVIVIPTSKGENRCWVRAIDVGYMCEDTITY ECPKLAVGNDPEDVDCWCDNQEVYVQYGRCTRTRHSKRSRRSINV SINVQTHGESSLVNKKEAWLDSTKATRYLMKTENWIIRNPGYAFL AAALGWMLGSNSGQRVVFTILLLLVAPAYSFNCLGMGNRDFIEGA SGATWVDLVLEGDSCLTIMANDKPTLDVRMINIEASQLAEVRSYC YHASINVTDISTVARCPTTGEAHNEKRADSSYVCKQGFTDRGWGN GCGLFGKGSIDTCAKFSCTNKAIGRMIQPENIKYEVGIFVHGTTT SENHGNYSAQVGASQAAKFTVTPNAPSITLKLGDYGEVTLDCEPR SGLNTEAFYVMTVGSKSFLVHREWFHDLSLPWTSPSSTAWRNREL LMEFEEAHATKQSINVVALGSQEGGLHQALAGAIVVEYSSSINVK LTSGHLKCRLKMDKLALKGTTYGMCTEKFSFAKNPADTGHGTVVI ELTYSGSDGPCKIPIVSINVASLNDMTPVGRLVTVNPFVATSSSN SKVLVEMEPPFGDSYIVVGRGDKQINHHWHKAGSTLGKAFSTTLK GAQRLAALGDTAWDFGSIGGVFNSIGKAVHQVFGGAFRTLFGGMS WITQGLMGALLLWMGVNARDRSIALAFLATGGVLVFLATNVHADT GCAIDITRKEMRCGSGIFVHNDVEAWVDRYKYLPETPRSLAKIVH KAHQEGVCGVRSINVTRLEHQMWESINVRDELNVLLKENAVDLSI NVVVNKPVGRYRSAPKRLSMTQEKFEMGWKAWGKSILFAPELANS TFVVDGPETKECPDERRAWNSMQIEDFGFGITSTRVWLKIREENT DECDGAIIGTAVKGHVAVHSDLSYWIESRLNDTWKLERAVFGEVK SCTWPETHTLWGDGVEESELIIPHTIAGPRSKHNRREGYKTQNQG PWDENGIVLDFDYCPGTKVTITEDCGKRGPSIRTTTDSGKLITDW CCRSCSLPPLRFRTENGCWYGMEIRPVRHDETTLVRSQVDAFNGE MIDPFQLGLLVMFLATQEVLRKRWTARLTIPAVLGALLVLMLGGI TYTDMARYVVLVAAAFAEANSGGDVLHLALIAVFKIQPSFLVMNM LSARWTNQENVVLVLGAAFFQLASINVDLQIGVHGILNAAAIAWM IVRAITFPTTSTVAMPVLALLTPGMRALYLDTYRIILLVIGICSL LQERRKTMAKKKGAVLLGLALTSTGWFSPTTIAAGLMVCNPNKKR GWPATEFLSAVGLMFAIVGGLAELDIESMSIPFMLAGLMAVSYVV SGKATDMWLDRAADISWEMEAAITGSSRRLDVKLDDDGDFHLIDD PGVPWKVWLLRMSCIGLAALTPWAIVPAAFGYWLTLKTTKRGGVF WDTPSPKPCLKGDTTTGVYRIMARGILGTYQAGVGVMYENVFHTL WHTTRGAAIMSGEGKLTPYWGSINVKEDRISYGGPWRFDRKWNGT DDVQVIVVEPGKPAVNIQTKPGVFRTPFGEIGAVSLDYPRGTSGS PILDSNGDIIGLYGNGVELGDGSYVSAIVQGDRQEEPVPDAYTPG MLKKRQMTVLDLHPGSGKTRKILPQIIKDAIQQRLRTAVLAPTRV VAAEMAEALKGLPVRYQTSAVQREHQGNEIVDVMCHATLTHRLMS PNRVPNYNLFVMDEAHFTDPASIAARGYIATKVELGEAAAIFMTA TPPGTTDPFPDSNAPIHDLQDEIPDRAWSSGYEWITDYAGKTVWF VASINVKMGNEIAMCLQRAGKKVIQLNRKSYDTEYPKCKNGDWDF VITTDISEMGANFGASRVIDCRKSINVKPTILEEGEGRVILGNPS PITSASAAQRRGRVGRNPNQVGDEYHYGGATSEDDSNLAHWTEAK IMLDNIHMPNGLVAQLYGPEREKAFTMDGEYRLRGEEKKNFLELL RTADLPVWLAYKVASNGIQYTDRKWCFDGPRTNAILEDNTEVEIV TRMGERKILKPRWLDARVYADHQALKWFKDFAAGKRSAVSFIEVL GRMPEHFMGKTREALDTMYLVATAEKGGKAHRMALEELPDALETI TLIVAITVMTGGFFLLMMQRKGIGKMGLGALVLTLATFFLWAAEV PGTKIAGTLLVALLLMVVLIPEPEKQRSQTDNQLAVFLICVLTVV GVVAANEYGMLEKTKADLKSMFGGRTQASGLTGLPSMALDLRPAT AWALYGGSTVVLTPLLKHLITSEYVTTSLASISSQAGSLFVLPRG VPFTDLDLTVGLVFLGCWGQITLTTFLTAMVLVTLHYGYMLPGWQ AEALRAAQRRTAAGIMKNAVVDGMVATDVPELERTTPLMQKKVGQ VLLIGVSINVAAFLVNPNVTTVREAGVLVTAATLTLWDNGASAVW NSTTATGLCHVMRGSYLAGGSIAWTLIKNADKPSLKRGRPGGRTL GEQWKEKLNAMSRDEFFKYRREAIIEVDRTEARRARRENNIVGGH PVSRGSAKLRWLVEKGFVSPIGKVIDLGCGRGGWSYYAATLKKVQ EVKGYTKGGAGHEEPMLMQSYGWNLVSLKSGVDVFYKPSEPSDTL FCDIGESSPSPEVEEQRTLRVLEMTSDWLHRGPREFCIKVLCPYM PKVIEKMEVLQRRFGGGLVRLPLSRNSNHEMYWVSGAAGNVVHAV NMTSQVLLGRMDRTVWRGPKYEEDVNLGSGTRAVGKGEVHSNQEK IRKRIQKLKEEFATTWHKDPEHPYRTWTYHGSYEVKATGSASSLV NGVVKLMSKPWDAIANVTTMAMTDTTPFGQQRVFKEKVDTKAPEP PAGVKEVLNETTNWLWAHLSREKRPRLCTKEEFIKKVNSNAALGA VFAEQNQWSTAREAVGDPLFWEMVNEERENHLRGECHTCIYNMMG KREKKPGEFGKAKGSRAIWFMWLGARYLEFEALGFLNEDHWLSRE NSGGGVEGSGVQKLGYILRDIAGKQGGKMYADDTAGWDTRITRTD LENEAKVLELLDGEHRMLARAIIELTYRHKVVKVMRPAAGGKTVM DVISREDQRGSGQVVTYALNTFTNIAVQLVRLMEAEGVIGPQHLE QLPRKNKIAVRTWLFENGEERVTRMAISGDDCVVKPLDDRFATAL HFLNAMSKVRKDIQEWKPSHGWHDWQQVPFCSNHFQEIVMKDGRS IVVPCRGQDELIGRARISPGAGWNVKDTACLAKAYAQMWLLLYFH RRDLRLMANAICSAVPVDWVPTGRTSWSIHSKGEWMTTEDMLQVW NRVWIEENEWMMDKTPITSWTDVPYVGKREDIWCGSLIGTRSRAT WAENVYAAINQVRAIIGKENYVDYMTSLRRYEDVLIQEDRVI >gi|5233922831| MTKKPGGPGKNRAINMLKRGLPRVFPLVGVKRVVMSLLDGRGPVR 27 dbj|BAN62837.1| FVLALITFFKFTALAPTKALLGRWRAVEKSINVAMKHLTSFKREL polyprotein GTLIDAVNKRGKKQNKRGGNESSIMWLASLAIVAACAGAMKLSNF QGKLLMTINNTDIADVIVIPTSKGENRCWVRAIDVGYMCEDTITY ECPKLAVGNDPEDVDCWCDNQEVYVQYGRCTRTRHSKRSRRSINV SINVQTHGESSLVNKKEAWLDSTKATRYLMKTENWIIRNPGYAFL AAALGWMLGSNSGQRVVFTILLLLVAPAYSFNCLGMGNRDFIEGA SGATWVDLVLEGDSCLTIMANDKPTLDVRMINIEASQLAEVRSYC YHASINVTDISTVARCPTTGEAHNEKRADSSYVCKQGFTDRGWGN GCGLFGKGSIDTCAKFSCTNKAIGRMIQPENIKYEVGIFVHGTTT SENHGNYSAQVGASQAAKFTVTPNAPSITLKLGDYGEVTLDCEPR SGLNTEAFYVMTVGSKSFLVHREWFHDLSLPWTSPSSTAWRNREL LMEFEEAHATKQSINVVALGSQEGGLHQALAGAIVVEYSSSINVK LTSGHLKCRLKMDKLALKGTTYGMCTEKFSFAKNPADTGHGTVVI ELTYSGSDGPCKIPIVSINVASLNDMTPVGRLVTVNPFVATSSSN SKVLVEMEPPFGDSYIVVGRGDKQINHHWHKAGSTLGKAFSTTLK GAQRLAALGDTAWDFGSIGGVFNSIGKAVHQVFGGAFRTLFGGMS WITQGLMGALLLWMGVNARDRSIALAFLATGGVLVFLATNVHADT GCAIDITRKEMRCGSGIFVHNDVEAWVDRYKYLPETPRSLAKIVH KAHQEGVCGVRSINVTRLEHQMWESINVRDELNVLLKENAVDLSI NVVVNKPVGRYRSAPKRLSMTQEKFEMGWKAWGKSILFAPELANS TFVVDGPETKECPDERRAWNSMQIEDFGFGITSTRVWLKIREENT DECDGAIIGTAVKGHVAVHSDLSYWIESRLNDTWKLERAVFGEVK SCTWPETHTLWGDGVEESELIIPHTIAGPRSKHNRREGYKTQNQG PWDENGIVLDFDYCPGTKVTITEDCGKRGPSIRTTTDSGKLITDW CCRSCSLPPLRFRTENGCWYGMEIRPVRHDETTLVRSQVDAFNGE MIDPFQLGLLVMFLATQEVLRKRWTARLTIPAVLGALLVLMLGGI TYTDLARYVVLVAAAFAEANSGGDVLHLALIAVFKIQPSFLVMNM LSARWTNQENVVLVLGAAFFQLASINVDLQIGVHGILNAAAIAWM IVRAITFPTTSTVAMPVLALLTPGMRALYLDTYRIILLVIGICSL LQERRKTMAKKKGAVLLGLALTSTGWFSPTTIAAGLMVCNPNKKR GWPATEFLSAVGLMFAIVGGLAELDIESMSIPFMLAGLMAVSYVV SGKATDMWLDRAADISWEMEAAITGSSRRLDVKLDDDGDFHLIDD PGVPWKVWLLRMSCIGLAALTPWAIVPAAFGYWLTLKTTKRGGVF WDTPSPKPCLKGDTTTGVYRIMARGILGTYQAGVGVMYENVFHTL WHTTRGAAIMSGEGKLTPYWGSINVKEDRISYGGPWRFDRKWNGT DDVQVIVVEPGKPAVNIQTKPGVFRTPFGEIGAVSLDYPRGTSGS PILDSNGDIIGLYGNGVELGDGSYVSAIVQGDRQEEPVPDAYTPS MLKKRQMTVLDLHPGSGKTRKILPQIIKDAIQQRLRTAVLAPTRV VAAEMAEALKGLPVRYQTSAVQREHQGNEIVDVMCHATLTHRLMS PNRVPNYNLFVMDEAHFTDPASIAARGYIATKVELGEAAAIFMTA TPPGTTDPFPDSNAPIHDLQDEIPDRAWSSGYEWITDYAGKTVWF VASINVKMGNEIAMCLQRAGKKVIQLNRKSYDTEYPKCKNGDWDF VITADISEMGANFGASRVIDCRKSINVKPTILEEGEGRVILGNPS PITSASAAQRRGRVGRNPNQVGDEYHYGGATSEDDSNLAHWTEAK IMLDNIHMPNGLVAQLYGPEREKAFTMDGEYRLRGEEKKNFLELL RTADLPVWLAYKVASNGIQYTDRKWCFDGPRTNAILEDNTEVEIV TRMGERKILKPRWLDARVYADHQALKWFKDFAAGKRSAVSFIEVL GRMPEHFMGKTREALDTMYLVATAEKGGKAHRMALEELPDALETI TLIVAITVMTGGFFLLMMQRKGIGKMGLGALVLTLATFFLWAAEV PGTKIAGTLLVALLLMVVLIPEPEKQRSQTDNQLAVFLICVLTVV GVVAANEYGMLEKTKADLKSMFGGRTQASGLTGLPSMALDLRPAT AWALYGGSTVVLTPLLKHLITSEYVTTSLASISSQAGSLFVLPRG VPFTDLDLTVGLVFLGCWGQITLTTFLTAMVLVTLHYGYMLPGWQ AEALRAAQRRTAAGIMKNAVVDGMVATDVPELERTTPLMQKKVGQ VLLIGVSINVAAFLVNPNVTTVREAGVLVTAATLTLWDNGASAVW NSTTATGLCHVMRGSYLAGGSIAWTLIKNADKPSLKRGRPGGRTL GEQWKEKLNAMSRDEFFKYRREAIIEVDRTEARRARRENNIVGGH PVSRGSAKLRWLVEKGFVSPIGKVIDLGCGRGGWSYYAATLKKVQ EVKGYTKGGAGHEEPMLMQSYGWNLVSLKSGVDVFYKPSEPSDTL FCDIGESSPSPEVEEQRTLRVLEMTSDWLHRGPREFCIKVLCPYM PKVIEKMEVLQRRFGGGLVRLPLSRNSNHEMYWVSGAAGNVVHAV NMTSQVLLGRMDRTVWRGPKYEEDVNLGSGTRAVGKGEVHSNQEK IRKRIQKLKEEFATTWHKDPEHPYRTWTYHGSYEVKATGSASSLV NGVVKLMSKPWDAIANVTTMAMTDTTPFGQQRVFKEKVDTKAPEP PAGVKEVLNETTNWLWAHLSREKRPRLCTKEEFIKKVNSNAALGA VFAEQNQWSTAREAVGDPLFWEMVNEERENHLRGECHTCIYNMMG KREKKPGEFGKAKGSRAIWFMWLGARYLEFEALGFLNEDHWLSRE NSGGGVEGSGVQKLGYILRDIAGKQGGKMYADDTAGWDTRITRTD LENEAKVLELLDGEHRMLARAIIELTYRHKVVKVMRPAAGGKTVM DVISREDQRGSGQVVTYALNTFTNIAVQLVRLMEAEGVIGPQHLE QLPRKNKIAVRTWLFENGEERVARMAISGDDCVVKPLDDRFATAL HFLNAMSKVRKDIQEWKPSHGWHDWQQVPFCSNHFQEIVMKDGRS IVVPCRGQDELIGRARISPGAGWNVKDTACLAKAYAQMWLLLYFH RRDLRLMANAICSAVPVDWVPTGRTSWSIHSKGEWMTTEDMLQVW NRVWIEENEWMMDKTPITSWTDVPYVGKREDIWCGSLIGTRSRAT WAENVYAAINQVRAIIGKENYVDYMTSLRRYEDVLIQEDRVI >gi|307826670| MTKKPGGPGKNRAINMLKRGLPRVFPLVGVKRVVMSLLDGRGPVR 28 gb|ADN94469.1| FVLALVAFFKFTALAPTKALLGRWRAVEKSINVAMKHLTSFKREL polyprotein GTLIDAVNKRGKKQNKRGGNESSIVWLASLAIVTACAGAMKLSNF QGKLLMTINNTDIADVIVIPTSKGENRCWVRAIDVGYMCEDTITY ECPKLAVGNDPEDVDCWCDNQEVFVQYGRCTRTRHSKRSRRSINV SINVQTHGESSLVNKKEAWLDSTKATRYLMKTENWIIRNPGYAFL AAALGWMLGSNSGQRVVFTILLLLVAPAYSFNCLGMGNRDFIEGA SGATWVDLVLEGDSCLTIMANDKPTLDVRMINIEASQLAEVRSYC YHASINVTDISTVARCPTTGEAHNEKRADSSYVCKQGFTDRGWGN GCGLFGKGSIDTCAKFSCTSKAIGRTIQPENIKYEVGVFVHGTTT SENHGNYSAQVGASQAAKFTVTPNAPSITLKLGDYGEVTLDCEPR SGLNTEAFYVMTVGSKSFLVHREWFHDLSLPWTSPSSTAWRNREL LMEFEEAHATKQSINVVALGSQEGGLHQALAGAIVVEYSSSINVK LTSGHLKCRLKMDKLALKGTTYGMCTEKFSFAKNPADTGHGTVVI ELTYSGSDGPCKIPIVSINVASLNDMTPVGRLVTVNPFVATSSSN SKVLVEMEPPFGDSYIVVGRGDKQINHHWHKAGSTLGKAFSTTLK GAQRLAALGDTAWDFGSIGGVFNSIGKAVHQVFGGAFRTLFGGMS WITQGLMGALLLWMGVNARDRSIALAFLATGGVLVFLATNVHADT GCAIDITRKEMRCGSGIFVHNDVEAWVDRYKYLPETPRSLAKIVH KAHQEGVCGVRSINVTRLEHQMWESINVRDELNVLLKENAVDLSI NVVVNKPVGRYRSAPKRLSMTQEKFEMGWKAWGKSILFAPELANS TFVVDGPETKECPDERRAWNSMQIEDFGFGITSTRVWLKIREEDT DECDGAIIGTAVKGHVAVHSDLSYWIESRFNDTWKLERAVFGEVK SCTWPETHTLWGDGVEESELIIPHTIAGPRSKHNRREGYKTQNQG PWDENGIVLDFDYCPGTKVTITEDCGKRGPSIRTTTDSGKLITDW CCRSCSLPPLRFRTENGCWYGMEIRPVRHDETTLVRSQVDAFNGE MIDPFQLGLLVMFLATQEVLRKRWTARLTIPAVLGALLVLMLGGI TYTDLARYVVLVAAAFAEANSGGDVLHLALIAVFKIQPAFLVMNM LSARWTNQENMVLVLGAAFFQLASINVDLQIGVHGILNAAAIAWM IVRAITFPTTSTVTMPVLALLTPGMRALYLDTYRIILLVIGICSL LQERRKTMAKKKGAVLLGLALTSTGWFSPTTIAAGLMVCNPNKKR GWPATEFLSAVGLMFAIVGGLAELDIESMSIPFMLAGLMAVSYVV SGKATDMWLDRAADISWEMEAAITGSSRRLDVKLDDDGDFHLIDD PGVPWKVWLLRMSCIGLAALTPWAIVPAAFGYWLTLKTTKRGGVF WDTPSPKPCLKGDTTTGVYRIMARGILGTYQAGVGVMYENVFHTL WHTTRGAAIMSGEGKLTPYWGSINVKEDRISYGGPWRFDRKWNGT DDVQVIVVEPGKPAVNIQTKPGVFRTPFGEIGAVSLDYPRGTSGS PILDSNGDIIGLYGNGVELGDGSYVSAIVQGDRQEEPVPDAYTPS MLKKRQMTVLDLHPGSGKTRKILPQIIKDAIQQRLRTAVLAPTRV VAAEMAEALRGLPVRYQTSAVQREHQGNEIVDVMCHATLTHRLMS PNRVPNYNLFVMDEAHFTDPASIAARGYIATKVELGEAAAIFMTA TPPGTTDPFPDSNAPIHDLQDEIPDRAWSSGYEWITEYAGKTVWF VASINVKMGNEIAMCLQRAGKKVIQLNRKSYDTEYPKCKNGDWDF VITTDISEMGANFGASRVIDCRKSINVKPTILEEGEGRVILGNPS PITSASAAQRRGRVGRNPNQVGDEYHYGGATSEDDSNLAHWTEAK IMLDNIHMPNGLVAQLYGPEREKAFTMDGEYRLRGEEKKNFLELL RTADLPVWLAYKVASNGIQYTDRKWCFDGPRTNAILEDNTEVEIV TRMGERKILKPRWLDARVYADHQALKWFKDFAAGKRSAVSFIEVL GRMPEHFMGKTREALDTMYLVATAEKGGKAHRMALEELPDALETI TLIVAITVMTGGFFLLMMQRKGIGKMGLGALVLTLATFFLWAAEV PGTKIAGTLLVALLLMVVLIPEPEKQRSQTDNQLAVFLICVLTVV GVVAANEYGMLEKTKADLKSMFGGRTQAPGLTGLPSMALDLRPAT AWALYGGSTVVLTPLLKHLITSEYVTTSLASISSQAGSLFVLPRG VPFTDLDLTVGLVFLGCWGQITLTTFLTAMVLVTLHYGYMLPGWQ AEALRAAQRRTAAGIMKNAVVDGMVATDVPELERTTPLMQKKVGQ VLLIGVSINVAAFLVNPNVTTVREAGVLVTAATLTLWDNGASAVW NSTTATGLCHVMRGSYLAGGSIAWTLIKNADKPSLKRGRPGGRTL GEQWKEKLNAMSRDEFFKYRREAIIEVDRTEARRARRENNIVGGH PVSRGSAKLRWLVEKGFVSPIGKVIDLGCGRGGWSYYAATLKKVQ EVKGYTKGGAGHEEPMLMQSYGWNLVSLKSGVDVFYKPSEPSDTL FCDIGESSPSPEVEEQRTLRVLEMTSDWLHRGPREFCIKVLCPYM PKVIEKMEVLQRRFGGGLVRLPLSRNSNHEMYWVSGAAGNVVHAV NMTSQVLLGRMDRTVWRGPKYEEDVNLGSGTRAVGKGEVHSNQEK IRKRIQKLREEFATTWHKDPEHPYRTWTYHGSYEVKATGSASSLV NGVVKLMSKPWDAIANVTTMAMTDTTPFGQQRVFKEKVDTKAPEP PAGVKEVLNETTNWLWAHLSREKRPRLCTKEEFIKKVNSNAALGA VFAEQNQWSTAREAVGDPLFWEMVDEERENHLRGECHTCIYNMMG KREKKPGEFGKAKGSRAIWFMWLGARYLEFEALGFLNEDHWLSRE NSGGGVEGSGVQKLGYILRDIAGKQGGKMYADDTAGWDTRITRTD LENEAKVLELLDGEHRMLARAIIELTYRHKVVKVMRPAAGGKTVM DVISREDQRGSGQVVTYALNTFTNIAVQLVRLMEAEGVIGPQHLE QLPRKNKIAVRTWLFENGEERVTRMAISGDDCVVKPLDDRFATAL HFLNAMSKVRKDIQEWKPSHGWHDWQQVPFCSNHFQEIVMKDGRS IVVPCRGQDELIGRARISPGAGWNVKDTACLAKAYAQMWLLLYFH RRDLRLMANAICSAVPVDWVPTGRTSWSIHSKGEWMTTEDMLQVW NRVWIEENEWMTDKTPITSWTDVPYVGKREDIWCGSLIGTRSRAT WAENIYAAINQVRAVIGKENYVDYMTSLRRYEDVLIQEDRVI >gi|307826668| MTKKPGGPGKNRAINMLKRGLPRVFPLVGVKRVVMSLLDGRGPVR 29 gb|ADN94468.1| FVLALITFFKFTALAPTKALLGRWRAVEKSINVAMKHLTSFKREL polyprotein GTLIDAVNKRGKKQNKRGGNESSIMWLASLAIVTACAGAMKLSNF QGKLLMTINNTDIADVIVIPTSKGENRCWVRAIDVGYMCEDTITY ECPKLAVGNDPEDVDCWCDNQEVFVQYGRCTRTRHSKRSRRSINV SINVQTHGESSLVNKKEAWLDSTKATRYLMKTENWIIRNPGYAFL AAALGWMLGSNSGQRVVFTILLLLVAPAYSFNCLGMGNRDFIEGA SGATWVDLVLEGDSCLTIMANDKPTLDVRMINIEASQLAEVRSYC YHASINVTDISTVARCPTTGEAHNEKRADSSYVCKQGFTDRGWGN GCGLFGKGSIDTCAKFSCTSKAIGRTIQPENIKYEVGVFVHGTTT SENHGNYSAQVGASQAAKFTVTPNAPSITLKLGDYGEVTLDCEPR SGLNTEAFYVMTVGSKSFLVHREWFHDLSLPWTSPSSTAWRNREL LMEFEEAHATKQSINVVALGSQEGGLHQALAGAIVVEYSSSINVK LTSGHLKCRLKMDKLALKGTTYGMCTEKFSFAKNPADTGHGTVVI ELTYSGSDGPCKIPIVSINVASLNDMTPVGRLVTVNPFVATSSSN SKVLVEMEPPFGDSYIVVGRGDKQINHHWHKAGSTLGKAFSTTLK GAQRLAALGDTAWDFGSIGGVFNSIGKAVHQVFGGAFRTLFGGMS WITQGLMGALLLWMGVNARDRSIALAFLATGGVLVFLATNVHADT GCAIDITRKEMRCGSGIFVHNDVEAWVDRYKYLPETPRSLAKIVH KAHQEGVCGVRSINVTRLEHQMWESINVRDELNVLLKENAVDLSI NVVVNKPVGRYRSAPKRLSMTQEKFEMGWKAWGKSILFAPELANS TFVVDGPETKECPDERRAWNSMQIEDFGFGITSTRVWLKIREEDT DECDGAIIGTAVKGHVAVHSDLSYWIESRLNDTWKLERAVFGEVK SCTWPETHTLWGDGVEESELIIPHTIAGPKSKHNRREGYKTQNQG PWDENGIVLDFDYCPGTKVTITEDCGKRGPSIRTTTDSGKLITDW CCRSCSLPPLRFRTENGCWYGMEIRPVRHDETTLVRSQVDAFNGE MIDPFQLGLLVMFLATQEVLRKRWTARLTIPAVLGALLVLMLGGI TYTDLARYVVLVAAAFAEANSGGDVLHLALIAVFKIQPAFLVMNM LSARWTNQENMVLVLGAAFFQLASINVDLQIGVHGILNAAAIAWM IVRAITFPTTSTVTMPVLALLTPGMRALYLDTYRIILLVIGTCSL LQERRKTMAKKKGAVLLGLALTSTGWFSPTTIAAGLMVCNPNKKR GWPATEFLSAVGLMFAIVGGLAELDIESMSIPFMLAGLMAVSYVV SGKATDMWLDRAADISWEMEAAITGSSRRLDVKLDDDGDFHLIDD PGVPWKVWLLRMSCIGLAALTPWAIVPAAFGYWLTLKTTKRGGVF WDTPSPKPCLKGDTTTGVYRIMARGILGTYQAGVGVMYENVFHTL WHTTRGAAIMSGEGKLTPYWGSINVKEDRISYGGPWRFDRKWNGT DDVQVIVVEPGKPAVNIQTKPGVFRTPFGEIGAVSLDYPRGTSGS PILDSNGDIIGLYGNGVELGDGSYVSAIVQGDRQEEPVPDAYTPS MLKKRQMTVLDLHPGSGKTRKILPQIIKDAIQQRLRTAVLAPTRV VAAEMAEALRGLPVRYQTSAVQREHQGNEIVDVMCHATLTHRLMS PNRVPNYNLFVMDEAHFTDPASIAARGYIATKVELGEAAAIFMTA TPPGTTDPFPDSNAPIHDLQDEIPDRAWSSGYEWITEYAGKTVWF VASINVKMGNEIAMCLQRAGKKVIQLNRKSYDTEYPKCKNGDWDF VITTDISEMGANFGASRVIDCRKSINVKPTILEEGEGRVILGNPS PITSASAAQRRGRVGRNPNQVGDEYHYGGATSEDDSNLAHWTEAK IMLDNIHMPNGLVAQLYGPEREKAFTMDGEYRLRGEEKKNFLELL RTADLPVWLAYKVASNGIQYTDRKWCFDGPRTNAILEDNTEVEIV TRMGERKILKPRWLDARVYADHQALKWFKDFAAGKRSAVSFIEVL GRMPEHFMGKTREALDTMYLVATAEKGGKAHRMALEELPDALETI TLIVAITVMTGGFFLLMMQRKGIGKMGLGALVLTLATFFLWAAEV PGTKIAGTLLVALLLMVVLIPEPEKQRSQTDNQLAVFLICVLTVV GVVAANEYGMLEKTKADLKSMFGGRTQAPGLTGLPSMALDLRPAT AWALYGGSTVVLTPLLKHLITSEYVTTSLASISSQAGSLFVLPRG VPFTDLDLTVGLVFLGCWGQITLTTFLTAMVLVTLHYGYMLPGWQ AEALRAAQRRTAAGIMKNAVVDGMVATDVPELERTTPLMQKKVGQ VLLIGVSINVAAFLVNPNVTTVREAGVLVTAATLTLWDNGASAVW NSTTATGLCHVMRGSYLAGGSIAWTLIKNADKPSLKRGRPGGRTL GEQWKEKLNAMSRDEFFKYRREAIIEVDRTEARRARRENNIVGGH PVSRGSAKLRWLVEKGFVSPIGKVIDLGCGRGGWSYYAATLKKVQ EVKGYTKGGAGHEEPMLMQSYGWNLVSLKSGVDVFYKPSEPSDTL FCDIGESSPSPEVEEQRTLRVLEMTSDWLHRGPREFCIKVLCPYM PKVIEKMEVLQRRFGGGLVRLPLSRNSNHEMYWVSGAAGNVVHAV NMTSQVLLGRMDRTVWRGPKYEEDVNLGSGTRAVGKGEVHSNQEK IRKRIQKLREEFATTWHKDPEHPYRTWTYHGSYEVKATGSASSLV NGVVKLMSKPWDAIANVTTMAMTDTTPFGQQRVFKEKVDTKAPEP PAGVKEVLNETTNWLWAHLSREKRPRLCTKEEFIKKVNSNAALGA VFAEQNQWSTAREAVGDPLFWEMVDEERENHLRGECHTCIYNMMG KREKKPGEFGKAKGSRAIWFMWLGARYLEFEALGFLNEDHWLSRE NSGGGVEGSGVQKLGYILRDIAGKQGGKMYADDTAGWDTRITRTD LENEAKVLELLDGEHRMLARAIIELTYRHKVVKVMRPAAGGKTVM DVISREDQRGSGQVVTYALNTFTNIAVQLVRLMEAEGVIGPQHLE QLPRKNKIAVRTWLFENGEERVTRMAISGDDCVVKPLDDRFATAL HFLNAMSKVRKDIQEWKPSHGWHDWQQVPFCSNHFQEIVMKDGRS IVVPCRGQDELIGRARISPGAGWNVKDTACLAKAYAQMWLLLYFH RRDLRLMANAICSAVPVDWVPTGRTSWSIHSKGEWMTTEDMLQVW NRVWIEENEWMMDKTPITSWTDVPYVGKREDIWCGSLIGTRSRAT WAENIYAAINQVRAVIGKENYVDYMISLRRYEDVLIQEDRVI Underlined sequence corresponds to a signal peptide, which may be omitted from each sequence. Thus, any RNA vaccine provided herein may encode an antigen represented by a sequence of Table 5, with or without the underlined signal peptide.

TABLE 6 JEV strains/isolates, Envelope proteins/variants - Homo sapiens GenBank Name Accession polyprotein [Japanese encephalitis virus] AHC56284.1 polyprotein [Japanese encephalitis virus] AEO86789.1 polyprotein [Japanese encephalitis virus] AIN36637.1 polyprotein [Japanese encephalitis virus] BAD81041.1 polypeptide [Japanese encephalitis virus] AAQ73508.1 polyprotein [Japanese encephalitis virus] BAI99560.1 polyprotein [Japanese encephalitis virus] AAD20233.1 polyprotein [Japanese encephalitis virus] AEO86778.1 polypeptide [Japanese encephalitis virus] AAQ73507.1 polyprotein [Japanese encephalitis virus] AHK05344.1 polyprotein [Japanese encephalitis virus] AEO72437.1 hypothetical protein JEVgp1 [Japanese encephalitis virus] NP_059434.1 polyprotein [Japanese encephalitis virus] AGT38389.1 polyprotein [Japanese encephalitis virus] BAI99561.1 polyprotein [Japanese encephalitis virus] AEO72436.1 polyprotein [Japanese encephalitis virus] AGW82423.1 polyprotein [Japanese encephalitis virus] AGT17711.1 polyprotein [Japanese encephalitis virus] BAJ51953.2 polyprotein [Japanese encephalitis virus] BAI99562.1 polyprotein [Japanese encephalitis virus] AEO86792.1 polyprotein [Japanese encephalitis virus] BAJ51960.2 polyprotein [Japanese encephalitis virus] AEO72439.1 polyprotein [Japanese encephalitis virus] BAD81039.1 polypeptide [Japanese encephalitis virus] AAQ73511.1 polyprotein [Japanese encephalitis virus] AEO86784.1 polyprotein [Japanese encephalitis virus] AEO72438.1 polyprotein [Japanese encephalitis virus] AAT00231.1 polyprotein [Japanese encephalitis virus] BAJ51952.2 polyprotein [Japanese encephalitis virus] BAJ51954.2 polyprotein [Japanese encephalitis virus] BAJ51955.2 polypeptide [Japanese encephalitis virus] AAQ73510.1 polyprotein [Japanese encephalitis virus] ADN94471.1 polypeptide [Japanese encephalitis virus] AAQ73513.1 polyprotein [Japanese encephalitis virus] AAA21436.1 polyprotein [Japanese encephalitis virus] BAJ51951.2 polyprotein [Japanese encephalitis virus] AEO72440.1 polypeptide [Japanese encephalitis virus] AAQ73512.1 polyprotein [Japanese encephalitis virus] AAM27886.1 polyprotein [Japanese encephalitis virus] AAM27885.1 polyprotein [Japanese encephalitis virus] AEO72432.1 polyprotein [Japanese encephalitis virus] BAF02840.1 polyprotein [Japanese encephalitis virus] AAB66485.1 polyprotein [Japanese encephalitis virus] BAJ51958.2 polyprotein [Japanese encephalitis virus] BAJ51959.2 polyprotein [Japanese encephalitis virus] CCI69572.1 polyprotein [Japanese encephalitis virus] AAS79438.1 polyprotein [Japanese encephalitis virus] AEO72435.1 polyprotein [Japanese encephalitis virus] AEO72433.1 polypeptide [Japanese encephalitis virus] AAQ73514.1 polyprotein [Japanese encephalitis virus] ABQ18323.1 polypeptide [Japanese encephalitis virus] AAQ73509.1 polyprotein [Japanese encephalitis virus] AIN36636.1 polyprotein [Japanese encephalitis virus] BAD81040.1 polyprotein [Japanese encephalitis virus] AGL09208.1 polyprotein [Japanese encephalitis virus] AEO72421.1 polyprotein; Contains: Peptide 2k; Capsid protein C; Core protein prM; P27395.1 Peptide pr; Small envelope protein M; Matrix protein; Envelope protein E; Non-structural protein 1; Non-structural protein 2A, Serine protease subunit NS2B; Flavivirin protease NS2B regulatory subunit; Non-structural protein 2B; Serine protease NS3; Flavivirin protease NS3 catalytic subunit; Non- structural protein 3; Non-structural protein 4A; Non-structural protein 4B; Non-structural protein 5 polyprotein [Japanese encephalitis virus] AEO86794.1 polyprotein [Japanese encephalitis virus] ACN39715.1 polyprotein [Japanese encephalitis virus] AEO72429.1 polyprotein [Japanese encephalitis virus] AEO72427.1 polyprotein [Japanese encephalitis virus] AEO72426.1 polyprotein [Japanese encephalitis virus] AEY64199.1 polyprotein [Japanese encephalitis virus] AEO72434.1 polyprotein [Japanese encephalitis virus] AEO72431.1 polyprotein [Japanese encephalitis virus] AEO72428.1 polyprotein [Japanese encephalitis virus] AAS79437.1 polyprotein [Japanese encephalitis virus] AEO72424.1 polyprotein [Japanese encephalitis virus] AIN36640.1 polyprotein [Japanese encephalitis virus] ACN39716.1 polyprotein [Japanese encephalitis virus] ABL60896.1 polyprotein [Japanese encephalitis virus] AIN36635.1 polyprotein [Japanese encephalitis virus] ABQ52691.1 polyprotein [Japanese encephalitis virus] AFP33182.1 polyprotein [Japanese encephalitis virus] AEO86781.1 polyprotein [Japanese encephalitis virus] AFP33181.1 polyprotein [Japanese encephalitis virus] AEO72423.1 polyprotein [Japanese encephalitis virus] AEO72430.1 polyprotein [Japanese encephalitis virus] AEY64200.1 polyprotein [Japanese encephalitis virus] AIN36639.1 polyprotein [Japanese encephalitis virus] ABU94627.1 polyprotein [Japanese encephalitis virus] AEO86793.1 polyprotein [Japanese encephalitis virus] AEO72422.1 polyprotein [Japanese encephalitis virus] AAB66484.1 polyprotein [Japanese encephalitis virus] ABU94628.1 polyprotein [Japanese encephalitis virus] ABU94629.1 polyprotein [Japanese encephalitis virus] AAB18951.1 C, prM, M, E, NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5 AAA64561.1 polyprotein [Japanese encephalitis virus] polyprotein [Japanese encephalitis virus] ACX54354.2 polyprotein [Japanese encephalitis virus] AEO72420.1 polyprotein [Japanese encephalitis virus] AEO86785.1 polyprotein [Japanese encephalitis virus] AAF34187.1 polyprotein [Japanese encephalitis virus] AAC29474.1 polyprotein [Japanese encephalitis virus] AEO72425.1 polyprotein [Japanese encephalitis virus] AAM77650.2 polyprotein [Japanese encephalitis virus] AAF34186.1 polyprotein [Japanese encephalitis virus] AEO86782.1 polyprotein [Japanese encephalitis virus] AAL77444.1 polyprotein [Japanese encephalitis virus] AAL08020.1 polyprotein [Japanese encephalitis virus] AFC87613.1 Japanese encephalitis virus isolate 08P38 envelope protein gene, partial cds FJ943489.1 Japanese encephalitis virus isolate 08P37 envelope protein gene, partial cds FJ943488.1 Japanese encephalitis virus isolate 07P127 envelope protein gene, partial FJ943487.1 cds Japanese encephalitis virus isolate 05P75 envelope protein gene, partial cds FJ943480.1 Japanese encephalitis virus E gene for polyprotein, partial cds, strain: AB920448.1 JE/Sw/Chiba/150/2007 Japanese encephalitis virus E gene for polyprotein, partial cds, strain: AB920447.1 JE/Sw/Kumamoto/104/2006 Japanese encephalitis virus gene for polyprotein, envelope protein, partial AB538665.2 cds, strain: Mo/Toyama/2554c/2007 Japanese encephalitis virus gene for polyprotein, partial cds, strain: AB231464.2 JEV/sw/Kagawa/35/2004 Japanese encephalitis virus gene for polyprotein, complete cds, strain: AB920399.1 JEV/sw/Okinawa/127/2012 Japanese encephalitis virus E gene for polyprotein, partial cds, strain: AB898105.1 Sw/Kochi/25/2005 Japanese encephalitis virus isolate YN79Bao83, complete genome JN381851.1 Japanese encephalitis virus gene for polyprotein, envelope protein, partial AB538661.2 cds, strain: Mo/Toyama/2506c/2007 Japanese encephalitis virus isolate 07P83 envelope protein gene, partial cds FJ943485.1 Japanese encephalitis virus isolate SH03-127 envelope protein (E) gene, DQ404101.1 partial cds Japanese encephalitis virus gene for envelope protein, partial cds, strain: LC079039.1 JEV/sw/Okinawa/186/2014 Japanese encephalitis virus gene for envelope protein, partial cds, strain: LC075515.1 JEV/sw/Okinawa/153/2015 Japanese encephalitis virus strain SH-96 polyprotein gene, envelope protein AY555760.1 region, partial cds Japanese encephalitis virus strain JE-91 envelope protein gene, partial cds GQ415355.1 Japanese encephalitis virus strain SH-101 polyprotein gene, envelope AY555761.1 protein region, partial cds Japanese encephalitis virus isolate K96A07 E protein gene, partial cds FJ938219.1 Japanese encephalitis virus isolate SH03-129 envelope protein (E) gene, DQ404103.1 partial cds Japanese encephalitis virus strain SC09-X29 envelope protein (E) gene, JQ411672.1 partial cds Japanese encephalitis virus isolate SH53, complete genome JN381850.1 Japanese encephalitis virus isolate K94A07 E protein gene, partial cds FJ938216.1 Japanese encephalitis virus isolate SH03-124 envelope protein (E) gene, DQ404100.1 partial cds Japanese encephalitis virus isolate SH03-109 envelope protein (E) gene, DQ404098.1 partial cds Japanese encephalitis virus strain VN105/Viet Nam/2002/Mosquito AY376468.1 envelope protein (E) gene, partial cds Japanese encephalitis virus isolate 97P82 envelope protein gene, partial cds FJ943472.1 Japanese encephalitis virus isolate JaNAr32-04 E protein gene, partial cds FJ185151.1 Japanese encephalitis virus isolate JaNAr07-04 E protein gene, partial cds FJ185144.1 Japanese encephalitis virus isolate SH05-24 envelope protein (E) gene, DQ404108.1 partial cds Japanese encephalitis virus strain SH-53 polyprotein gene, envelope protein AY555757.1 region, partial cds Japanese encephalitis virus gene for polyprotein, partial cds, strain: AB174838.1 JEV/swine/Hiroshima/38/2000 Japanese encephalitis virus E gene for polyprotein, partial cds, strain: AB920445.1 JE/Sw/Kumamoto/65/2005 Japanese encephalitis virus isolate SH80, complete genome JN381848.1 Japanese encephalitis virus isolate 06P152 envelope protein gene, partial FJ943481.1 cds Japanese encephalitis virus gene for polyprotein, partial cds, strain: AB231465.1 JEV/sw/Hiroshima/25/2002 Japanese encephalitis virus isolate KL113/SD/CHN/13 envelope protein KJ190848.1 gene, partial cds Japanese encephalitis virus isolate ZJ09-108 envelope protein gene, partial JN216866.1 cds Japanese encephalitis virus isolate ZJ09-52 envelope protein gene, partial JN216865.1 cds Japanese encephalitis virus isolate SH03105, complete genome JN381846.1 Japanese encephalitis virus isolate HN0421, complete genome JN381841.1 Japanese encephalitis virus genomic RNA, complete genome, strain: AB594829.1 JEV/eq/Tottori/2003 Japanese encephalitis virus isolate K01-GN E protein gene, partial cds FJ938220.1 Japanese encephalitis virus isolate SH03-105 envelope protein (E) gene, DQ404097.1 partial cds Japanese encephalitis virus strain JaNAr0102/Japan/2002/Mosquito AY377577.1 envelope protein (E) gene, partial cds Japanese encephalitis virus strain SH-83 polyprotein gene, envelope protein AY555759.1 region, partial cds Japanese encephalitis virus strain SH-81 polyprotein gene, envelope protein AY555758.1 region, partial cds Japanese encephalitis virus gene for polyprotein, partial cds AB213007.1 Japanese encephalitis virus gene for polyprotein, partial cds, strain: AB112706.1 JEV/sw/Kagawa/24/2002 Japanese encephalitis virus strain JX61, complete genome GU556217.1 Japanese encephalitis virus strain JX67 polyprotein gene, partial cds FJ179365.1 Japanese encephalitis virus strain JX66 polyprotein gene, partial cds FJ179364.1 Japanese encephalitis virus E gene for polyprotein, partial cds, strain: AB920449.1 JE/Sw/Chiba/103/2008 Japanese encephalitis virus E gene for polyprotein, partial cds, strain: AB920446.1 JE/Sw/Kumamoto/81/2006 Japanese encephalitis virus isolate JEV/Taiwan/TPC0806c/M/2008 KF667316.1 polyprotein gene, complete cds Japanese encephalitis virus isolate LiC68/SD/CHN/10 envelope protein KJ190834.1 gene, partial cds Japanese encephalitis virus strain LN0702 envelope protein gene, partial cds JQ937353.1 Japanese encephalitis virus strain BL0653 envelope protein gene, partial cds JQ937352.1 Japanese encephalitis virus isolate K10CT621 polyprotein mRNA, partial JX018158.1 cds Japanese encephalitis virus isolate ZJ10-10 envelope protein gene, partial JN216868.1 cds Japanese encephalitis virus isolate LN02-102, complete genome JF706278.1 Japanese encephalitis virus isolate BL06-50, complete genome JF706270.1 Japanese encephalitis virus isolate SH03103, complete genome JN381847.1 Japanese encephalitis virus isolate GSBY0816, complete genome JN381842.1 Japanese encephalitis virus isolate XP174M-08 envelope protein gene, HM204527.1 partial cds Japanese encephalitis virus strain TPC0806c envelope protein gene, partial GQ260635.1 cds Japanese encephalitis virus isolate 03P145 envelope protein gene, partial FJ943478.1 cds Japanese encephalitis virus isolate 03P120 envelope protein gene, partial FJ943476.1 cds Japanese encephalitis virus isolate YN86-86266 envelope protein (E) gene, DQ404134.1 partial cds Japanese encephalitis virus isolate LN02-104 envelope protein (E) gene, DQ404086.1 partial cds Japanese encephalitis virus strain VN88/Viet Nam/2001/Swine blood AY376464.1 envelope protein (E) gene, partial cds Japanese encephalitis virus strain KV1899, complete genome AY316157.1 Japanese encephalitis virus KV1899 polyprotein mRNA, partial cds AF474075.1 Japanese encephalitis virus K94P05 envelope protein (E) gene, partial cds U34929.1 Japanese encephalitis virus isolate YN79-Bao83 envelope protein (E) gene, DQ404128.1 partial cds Japanese encephalitis virus strain JX0939 envelope protein gene, partial cds JQ937355.1 Japanese encephalitis virus strain SDJN0908 envelope protein gene, partial JQ937344.1 cds Japanese encephalitis virus strain HBZG0809 envelope protein gene, partial JQ937333.1 cds Japanese encephalitis virus isolate 1XG014 envelope protein gene, partial JX514947.1 cds Japanese encephalitis virus strain SC09-A38 envelope protein (E) gene, JQ411670.1 partial cds Japanese encephalitis virus isolate TC2010-5 envelope protein gene, partial JF499814.1 cds Japanese encephalitis virus isolate TC2009-3 envelope protein gene, partial JF499793.1 cds Japanese encephalitis virus isolate TC2009-3, complete genome JF499788.1 Japanese encephalitis virus isolate ZJ10-45 envelope protein gene, partial JN216870.1 cds Japanese encephalitis virus strain A10.881 polyprotein gene, partial cds JN587260.1 Japanese encephalitis virus isolate HN0621, complete genome JN381830.1 Japanese encephalitis virus isolate K05-GS E protein gene, partial cds FJ938223.1 Japanese encephalitis virus isolate 03P126 envelope protein gene, partial FJ943477.1 cds Japanese encephalitis virus isolate 03P113 envelope protein gene, partial FJ943475.1 cds Japanese encephalitis virus isolate SH03-128 envelope protein (E) gene, DQ404102.1 partial cds Japanese encephalitis virus isolate SC04-16 envelope protein (E) gene, DQ404092.1 partial cds Japanese encephalitis virus strain 95-167/Japan/1995/Swine blood envelope AY377579.1 protein (E) gene, partial cds Japanese encephalitis virus strain K94P05, complete genome AF045551.2 Japanese encephalitis virus gene for polyprotein, envelope protein, partial AB538609.2 cds, strain: Mo/Toyama/1155v/2005 Japanese encephalitis virus isolate YN83-Meng83-54 envelope protein (E) DQ404130.1 gene, partial cds Japanese encephalitis virus isolate SH-80 polyprotein gene, partial cds AY243841.1 Japanese encephalitis virus strain ZJ13-11 envelope protein gene, partial cds KJ000037.1 Japanese encephalitis virus strain LN0828 envelope protein gene, partial cds JQ937354.1 Japanese encephalitis virus isolate 3XG001 envelope protein gene, partial JX514948.1 cds

TABLE 7 Example 18 Test Conditions Dosage Dose 2^(nd) mRNA Conc. Volume + G# Antigen Route N = (ug) Vol (ul) 1^(st) dose dose LNP (mg/ml) Overage 1 SE_West Nile PRM-E IM 5 10 50 Day 0 Day 28 MC3 0.2 2 × 800 ul 2 SE_WNV_TX8559_EZE1_SP IM 5 10 50 Day 0 Day 28 MC3 0.2 2 × 800 ul 3 JEV-JEV_FL_PrM-E IM 5 10 50 Day 0 Day 28 MC3 0.2 2 × 800 ul 4 JEV_FL_E IM 5 10 50 Day 0 Day 28 MC3 0.2 2 × 800 ul 5 SE_EEEv_FL93-939_EZE1_SP IM 5 10 50 Day 0 Day 28 MC3 0.2 2 × 800 ul 6 SE_SINV_AR339- IM 5 10 50 Day 0 Day 28 MC3 0.2 2 × 800 ul BeAr436087_EZE1_SP 7 Bivalent-WN PRM-E + IM 5 10 50 Day 0 Day 28 MC3 0.2 2 × 800 ul JEV-JEV_FL_PrM-E 8 Trivalent: WN PRM-E + JEV-PrM-E + SIN IM 5 10 50 Day 0 Day28 MC3 0.2 1 × 800 ul 9 Tetra valent: WN PRM-E + JEV + SIN + IM 5 10 50 Day 0 Day28 MC3 0.2 1 × 800 ul EEE 10 PBS IM 5 50 Day 0 Day28 1 × 800 ul

TABLE 8 Example 18 Test Conditions Dosage Dose 2^(nd) mRNA Conc. Volume + G# Antigen Route N = (ug) Vol (ul) 1^(st) dose dose LNP (mg/ml) Overage 1 SE_West Nile PRM-E IM 5 10 50 Day 0 Day 28 MC3 0.2 2 × 800 ul 2 SE_WNV_TX8559_EZE1_SP IM 5 10 50 Day 0 Day 28 MC3 0.2 2 × 800 ul 3 JEV-JEV_FL_PrM-E IM 5 10 50 Day 0 Day 28 MC3 0.2 2 × 800 ul 4 JEV_FL_E IM 5 10 50 Day 0 Day 28 MC3 0.2 2 × 800 ul 5 Bivalent-WN PrM-E + IM 5 10 50 Day 0 Day 28 MC3 0.2 2 × 800 ul JEV-JEV_FL_PrM-E 6 Bivalent: SE_WNV_TX8559_EZE1_SP + IM 5 10 50 Day 0 Day28 MC3 0.2 1 × 800 ul JEV_FL_E 7 PBS IM 5 50 Day 0 Day28 1 × 800 ul

TABLE 9 WNV Nucleic Acid Sequences Antigen Nucleic Acid Sequence SEQ ID NO: WNV ORF: 30 SE_West Nile ATGTGGCTTGTCAGCTTGGCAATCGTAACAGCTTGTGCCGGAGCTAT PRM-E novel GAAGCTCTCTAACTTCCAAGGGAAGGTGATGATGACGGTAAATGCTA antigen CTGACGTCACAGATGTCATCACGATTCCAACAGCTGCTGGAAAGAAC CTATGCATTGTCAGAGCAATGGATGTGGGATACATGTGCGATGATAC TATCACTTATGAATGCCCAGTGCTGTCGGCTGGCAATGATCCAGAAG ACATCGACTGTTGGTGCACAAAGTCAGCAGTCTACGTCAGGTATGGA AGATGCACCAAGACACGCCACTCAAGACGCAGTCGGAGGTCACTGAC AGTGCAGACACACGGAGAAAGCACTCTAGCGAACAAGAAGGGGGCTT GGATGGACAGCACCAAGGCCACAAGGTATTTGGTAAAAACAGAATCA TGGATCTTGAGGAACCCTGGATATGCCCTGGTGGCAGCCGTCATTGG TTGGATGCTTGGGAGCAACACCATGCAGAGAGTTGTGTTTGTCGTGC TATTGCTTTTGGTGGCCCCAGCTTACAGCTTCAACTGCCTTGGAATG AGCAACAGAGACTTCTTGGAAGGAGTGTCTGGAGCAACATGGGTGGA TTTGGTTCTCGAAGGCGACAGCTGCGTGACTATCATGTCTAAGGACA AGCCTACCATCGATGTGAAGATGATGAATATGGAGGCGGCCAACCTG GCAGAGGTCCGCAGTTATTGCTATTTGGCTACCGTCAGCGATCTCTC CACCAAAGCTGCGTGCCCGACCATGGGAGAAGCTCACAATGACAAAC GTGCTGACCCAGCTTTTGTGTGCAGACAAGGAGTGGTGGACAGGGGC TGGGGCAACGGCTGCGGACTATTTGGCAAAGGAAGCATTGACACATG CGCCAAATTTGCCTGCTCTACCAAGGCAATAGGAAGAACCATCTTGA AAGAGAATATCAAGTATGAAGTGGCCATTTTTGTCCATGGGCCAACT ACTGTGGAGTCGCACGGAAACTACTCCACACAGGCTGGAGCCACTCA GGCAGGGAGATTCAGCATCACTCCTGCGGCGCCTTCATACACACTAA AGCTTGGAGAATATGGAGAGGTGACAGTGGACTGTGAACCACGGTCA GGGATTGACACCAATGCATACTACGTGATGACTGTTGGAACAAAGAC GTTCTTGGTCCATCGTGAGTGGTTCATGGACCTCAACCTCCCTTGGA GCAGTGCTGGAAGTACTGTGTGGAGGAACAGAGAGACGTTAATGGAG TTTGAGGAACCGCACGCCACGAAGCAGTCTGTGATAGCATTGGGCTC ACAAGAGGGAGCTCTGCATCAAGCTTTGGCTGGAGCCATTCCTGTGG AATTTTCAAGCAACACTGTCAAGTTGACGTCGGGTCATTTGAAGTGT AGAGTGAAGATGGAAAAATTGCAGTTGAAGGGAACAACCTATGGCGT CTGTTCAAAGGCTTTCAAGTTTCTTGGGACTCCCGCAGACACAGGTC ACGGCACTGTGGTGTTGGAATTGCAGTACACTGGCACGGACGGACCT TGCAAAGTTCCTATCTCGTCAGTGGCTTCATTGAACGACCTAACGCC AGTGGGTAGATTGGTCACTGTCAACCCTTTTGTTTCAGTGGCCACGG CCAACGCTAAGGTCCTGATTGAATTGGAACCACCCTTTGGAGACTCA TACATAGTGGTGGGCAGAGGAGAACAACAGATCAATCATCATTGGCA CAAGTCTGGAAGCAGCATTGGCAAAGCCTTTACAACCACCCTCAAAG GAGCGCAGAGACTAGCCGCTCTAGGAGACACAGCTTGGGACTTTGGA TCAGTTGGAGGGGTGTTCACCTCAGTTGGGAAGGCTGTCCATCAAGT GTTCGGAGGAGCATTCCGCTCACTGTTCGGAGGCATGTCCTGGATAA CGCAAGGATTGCTGGGGGCTCTCTTGTTGTGGATGGGCATCAATGCT CGTGATAGGTCCATAGCTCTCACGTTTCTCGCAGTTGGAGGAGTTCT GCTCTTCCTCTCCGTGAACGTGCATGCT SE_WNV_TX8 ORF: 31 559_E2E1 ATGTTCAACTGCCTTGGAATGAGCAACAGAGACTTCTTGGAAGGAGT WNV, USA GTCTGGAGCAACATGGGTGGATTTGGTTCTCGAAGGCGACAGCTGCG isolate TGACTATCATGTCTAAGGACAAGCCTACCATCGATGTGAAGATGATG AATATGGAGGCGGCCAACCTGGCAGAGGTCCGCAGTTATTGCTATTT GGCTACCGTCAGCGATCTCTCCACCAAAGCTGCGTGCCCGACCATGG GAGAAGCTCACAATGACAAACGTGCTGACCCAGCTTTTGTGTGCAGA CAAGGAGTGGTGGACAGGGGCTGGGGCAACGGCTGCGGACTATTTGG CAAAGGAAGCATTGACACATGCGCCAAATTTGCCTGCTCTACCAAGG CAATAGGAAGAACCATCTTGAAAGAGAATATCAAGTATGAAGTGGCC ATTTTTGTCCATGGGCCAACTACTGTGGAGTCGCACGGAAACTACTC CACACAGGCTGGAGCCACTCAGGCAGGGAGATTCAGCATCACTCCTG CGGCGCCTTCATACACACTAAAGCTTGGAGAATATGGAGAGGTGACA GTGGACTGTGAACCACGGTCAGGGATTGACACCAATGCATACTACGT GATGACTGTTGGAACAAAGACGTTCTTGGTCCATCGTGAGTGGTTCA TGGACCTCAACCTCCCTTGGAGCAGTGCTGGAAGTACTGTGTGGAGG AACAGAGAGACGTTAATGGAGTTTGAGGAACCGCACGCCACGAAGCA GTCTGTGATAGCATTGGGCTCACAAGAGGGAGCTCTGCATCAAGCTT TGGCTGGAGCCATTCCTGTGGAATTTTCAAGCAACACTGTCAAGTTG ACGTCGGGTCATTTGAAGTGTAGAGTGAAGATGGAAAAATTGCAGTT GAAGGGAACAACCTATGGCGTCTGTTCAAAGGCTTTCAAGTTTCTTG GGACTCCCGCAGACACAGGTCACGGCACTGTGGTGTTGGAATTGCAG TACACTGGCACGGACGGACCTTGCAAAGTTCCTATCTCGTCAGTGGC TTCATTGAACGACCTAACGCCAGTGGGTAGATTGGTCACTGTCAACC CTTTTGTTTCAGTGGCCACGGCCAACGCTAAGGTCCTGATTGAATTG GAACCACCCTTTGGAGACTCATACATAGTGGTGGGCAGAGGAGAACA ACAGATCAATCATCATTGGCACAAGTCTGGAAGCAGCATTGGCAAAG CCTTTACAACCACCCTCAAAGGAGCGCAGAGACTAGCCGCTCTAGGA GACACAGCTTGGGACTTTGGATCAGTTGGAGGGGTGTTCACCTCAGT TGGGAAGGCTGTCCATCAAGTGTTCGGAGGAGCATTCCGCTCACTGT TCGGAGGCATGTCCTGGATAACGCAAGGATTGCTGGGGGCTCTCTTG TTGTGGATGGGCATCAATGCTCGTGATAGGTCCATAGCTCTCACGTT TCTCGCAGTTGGAGGAGTTCTGCTCTTCCTCTCCGTGAACGTGCATG CT SE_WNV_TX8 mRNA sequence: 32 559_E2E1 G*GGGAAATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCA WNV, USA CCATGTTCAACTGCCTTGGAATGAGCAACAGAGACTTCTTGGAAGGA isolate GTGTCTGGAGCAACATGGGTGGATTTGGTTCTCGAAGGCGACAGCTG CGTGACTATCATGTCTAAGGACAAGCCTACCATCGATGTGAAGATGA TGAATATGGAGGCGGCCAACCTGGCAGAGGTCCGCAGTTATTGCTAT TTGGCTACCGTCAGCGATCTCTCCACCAAAGCTGCGTGCCCGACCAT GGGAGAAGCTCACAATGACAAACGTGCTGACCCAGCTTTTGTGTGCA GACAAGGAGTGGTGGACAGGGGCTGGGGCAACGGCTGCGGACTATTT GGCAAAGGAAGCATTGACACATGCGCCAAATTTGCCTGCTCTACCAA GGCAATAGGAAGAACCATCTTGAAAGAGAATATCAAGTATGAAGTGG CCATTTTTGTCCATGGGCCAACTACTGTGGAGTCGCACGGAAACTAC TCCACACAGGCTGGAGCCACTCAGGCAGGGAGATTCAGCATCACTCC TGCGGCGCCTTCATACACACTAAAGCTTGGAGAATATGGAGAGGTGA CAGTGGACTGTGAACCACGGTCAGGGATTGACACCAATGCATACTAC GTGATGACTGTTGGAACAAAGACGTTCTTGGTCCATCGTGAGTGGTT CATGGACCTCAACCTCCCTTGGAGCAGTGCTGGAAGTACTGTGTGGA GGAACAGAGAGACGTTAATGGAGTTTGAGGAACCGCACGCCACGAAG CAGTCTGTGATAGCATTGGGCTCACAAGAGGGAGCTCTGCATCAAGC TTTGGCTGGAGCCATTCCTGTGGAATTTTCAAGCAACACTGTCAAGT TGACGTCGGGTCATTTGAAGTGTAGAGTGAAGATGGAAAAATTGCAG TTGAAGGGAACAACCTATGGCGTCTGTTCAAAGGCTTTCAAGTTTCT TGGGACTCCCGCAGACACAGGTCACGGCACTGTGGTGTTGGAATTGC AGTACACTGGCACGGACGGACCTTGCAAAGTTCCTATCTCGTCAGTG GCTTCATTGAACGACCTAACGCCAGTGGGTAGATTGGTCACTGTCAA CCCTTTTGTTTCAGTGGCCACGGCCAACGCTAAGGTCCTGATTGAAT TGGAACCACCCTTTGGAGACTCATACATAGTGGTGGGCAGAGGAGAA CAACAGATCAATCATCATTGGCACAAGTCTGGAAGCAGCATTGGCAA AGCCTTTACAACCACCCTCAAAGGAGCGCAGAGACTAGCCGCTCTAG GAGACACAGCTTGGGACTTTGGATCAGTTGGAGGGGTGTTCACCTCA GTTGGGAAGGCTGTCCATCAAGTGTTCGGAGGAGCATTCCGCTCACT GTTCGGAGGCATGTCCTGGATAACGCAAGGATTGCTGGGGGCTCTCT TGTTGTGGATGGGCATCAATGCTCGTGATAGGTCCATAGCTCTCACG TTTCTCGCAGTTGGAGGAGTTCTGCTCTTCCTCTCCGTGAACGTGCA TGCTTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTT GGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCGT GGTCTTTGAATAAAGTCTGAGTGGGCGGCAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAATCTAG SE_WNV_TX8 ORF: 33 559_E2E1_SP ATGCAGAGAGTTGTGTTTGTCGTGCTATTGCTTTTGGTGGCCCCAGC WNV, USA TTACAGCTTCAACTGCCTTGGAATGAGCAACAGAGACTTCTTGGAAG isolate GAGTGTCTGGAGCAACATGGGTGGATTTGGTTCTCGAAGGCGACAGC TGCGTGACTATCATGTCTAAGGACAAGCCTACCATCGATGTGAAGAT GATGAATATGGAGGCGGCCAACCTGGCAGAGGTCCGCAGTTATTGCT ATTTGGCTACCGTCAGCGATCTCTCCACCAAAGCTGCGTGCCCGACC ATGGGAGAAGCTCACAATGACAAACGTGCTGACCCAGCTTTTGTGTG CAGACAAGGAGTGGTGGACAGGGGCTGGGGCAACGGCTGCGGACTAT TTGGCAAAGGAAGCATTGACACATGCGCCAAATTTGCCTGCTCTACC AAGGCAATAGGAAGAACCATCTTGAAAGAGAATATCAAGTATGAAGT GGCCATTTTTGTCCATGGGCCAACTACTGTGGAGTCGCACGGAAACT ACTCCACACAGGCTGGAGCCACTCAGGCAGGGAGATTCAGCATCACT CCTGCGGCGCCTTCATACACACTAAAGCTTGGAGAATATGGAGAGGT GACAGTGGACTGTGAACCACGGTCAGGGATTGACACCAATGCATACT ACGTGATGACTGTTGGAACAAAGACGTTCTTGGTCCATCGTGAGTGG TTCATGGACCTCAACCTCCCTTGGAGCAGTGCTGGAAGTACTGTGTG GAGGAACAGAGAGACGTTAATGGAGTTTGAGGAACCGCACGCCACGA AGCAGTCTGTGATAGCATTGGGCTCACAAGAGGGAGCTCTGCATCAA GCTTTGGCTGGAGCCATTCCTGTGGAATTTTCAAGCAACACTGTCAA GTTGACGTCGGGTCATTTGAAGTGTAGAGTGAAGATGGAAAAATTGC AGTTGAAGGGAACAACCTATGGCGTCTGTTCAAAGGCTTTCAAGTTT CTTGGGACTCCCGCAGACACAGGTCACGGCACTGTGGTGTTGGAATT GCAGTACACTGGCACGGACGGACCTTGCAAAGTTCCTATCTCGTCAG TGGCTTCATTGAACGACCTAACGCCAGTGGGTAGATTGGTCACTGTC AACCCTTTTGTTTCAGTGGCCACGGCCAACGCTAAGGTCCTGATTGA ATTGGAACCACCCTTTGGAGACTCATACATAGTGGTGGGCAGAGGAG AACAACAGATCAATCATCATTGGCACAAGTCTGGAAGCAGCATTGGC AAAGCCTTTACAACCACCCTCAAAGGAGCGCAGAGACTAGCCGCTCT AGGAGACACAGCTTGGGACTTTGGATCAGTTGGAGGGGTGTTCACCT CAGTTGGGAAGGCTGTCCATCAAGTGTTCGGAGGAGCATTCCGCTCA CTGTTCGGAGGCATGTCCTGGATAACGCAAGGATTGCTGGGGGCTCT CTTGTTGTGGATGGGCATCAATGCTCGTGATAGGTCCATAGCTCTCA CGTTTCTCGCAGTTGGAGGAGTTCTGCTCTTCCTCTCCGTGAACGTG CATGCT SE_WNV_TX8 mRNA sequence: 34 559_E2E1_SP G*GGGAAATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCA WNV, USA CCATGCAGAGAGTTGTGTTTGTCGTGCTATTGCTTTTGGTGGCCCCA isolate GCTTACAGCTTCAACTGCCTTGGAATGAGCAACAGAGACTTCTTGGA AGGAGTGTCTGGAGCAACATGGGTGGATTTGGTTCTCGAAGGCGACA GCTGCGTGACTATCATGTCTAAGGACAAGCCTACCATCGATGTGAAG ATGATGAATATGGAGGCGGCCAACCTGGCAGAGGTCCGCAGTTATTG CTATTTGGCTACCGTCAGCGATCTCTCCACCAAAGCTGCGTGCCCGA CCATGGGAGAAGCTCACAATGACAAACGTGCTGACCCAGCTTTTGTG TGCAGACAAGGAGTGGTGGACAGGGGCTGGGGCAACGGCTGCGGACT ATTTGGCAAAGGAAGCATTGACACATGCGCCAAATTTGCCTGCTCTA CCAAGGCAATAGGAAGAACCATCTTGAAAGAGAATATCAAGTATGAA GTGGCCATTTTTGTCCATGGGCCAACTACTGTGGAGTCGCACGGAAA CTACTCCACACAGGCTGGAGCCACTCAGGCAGGGAGATTCAGCATCA CTCCTGCGGCGCCTTCATACACACTAAAGCTTGGAGAATATGGAGAG GTGACAGTGGACTGTGAACCACGGTCAGGGATTGACACCAATGCATA CTACGTGATGACTGTTGGAACAAAGACGTTCTTGGTCCATCGTGAGT GGTTCATGGACCTCAACCTCCCTTGGAGCAGTGCTGGAAGTACTGTG TGGAGGAACAGAGAGACGTTAATGGAGTTTGAGGAACCGCACGCCAC GAAGCAGTCTGTGATAGCATTGGGCTCACAAGAGGGAGCTCTGCATC AAGCTTTGGCTGGAGCCATTCCTGTGGAATTTTCAAGCAACACTGTC AAGTTGACGTCGGGTCATTTGAAGTGTAGAGTGAAGATGGAAAAATT GCAGTTGAAGGGAACAACCTATGGCGTCTGTTCAAAGGCTTTCAAGT TTCTTGGGACTCCCGCAGACACAGGTCACGGCACTGTGGTGTTGGAA TTGCAGTACACTGGCACGGACGGACCTTGCAAAGTTCCTATCTCGTC AGTGGCTTCATTGAACGACCTAACGCCAGTGGGTAGATTGGTCACTG TCAACCCTTTTGTTTCAGTGGCCACGGCCAACGCTAAGGTCCTGATT GAATTGGAACCACCCTTTGGAGACTCATACATAGTGGTGGGCAGAGG AGAACAACAGATCAATCATCATTGGCACAAGTCTGGAAGCAGCATTG GCAAAGCCTTTACAACCACCCTCAAAGGAGCGCAGAGACTAGCCGCT CTAGGAGACACAGCTTGGGACTTTGGATCAGTTGGAGGGGTGTTCAC CTCAGTTGGGAAGGCTGTCCATCAAGTGTTCGGAGGAGCATTCCGCT CACTGTTCGGAGGCATGTCCTGGATAACGCAAGGATTGCTGGGGGCT CTCTTGTTGTGGATGGGCATCAATGCTCGTGATAGGTCCATAGCTCT CACGTTTCTCGCAGTTGGAGGAGTTCTGCTCTTCCTCTCCGTGAACG TGCATGCTTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCC CCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCC CCGTGGTCTTTGAATAAAGTCTGAGTGGGCGGCAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAATCTAG WNV mRNA Sequences WNV ORF: 35 SE_West Nile AUGUGGCUUGUCAGCUUGGCAAUCGUAACAGCUUGUGCCGGAGCUAU PRM-E novel GAAGCUCUCUAACUUCCAAGGGAAGGUGAUGAUGACGGUAAAUGCUA antigen CUGACGUCACAGAUGUCAUCACGAUUCCAACAGCUGCUGGAAAGAAC CUAUGCAUUGUCAGAGCAAUGGAUGUGGGAUACAUGUGCGAUGAUAC UAUCACUUAUGAAUGCCCAGUGCUGUCGGCUGGCAAUGAUCCAGAAG ACAUCGACUGUUGGUGCACAAAGUCAGCAGUCUACGUCAGGUAUGGA AGAUGCACCAAGACACGCCACUCAAGACGCAGUCGGAGGUCACUGAC AGUGCAGACACACGGAGAAAGCACUCUAGCGAACAAGAAGGGGGCUU GGAUGGACAGCACCAAGGCCACAAGGUAUUUGGUAAAAACAGAAUCA UGGAUCUUGAGGAACCCUGGAUAUGCCCUGGUGGCAGCCGUCAUUGG UUGGAUGCUUGGGAGCAACACCAUGCAGAGAGUUGUGUUUGUCGUGC UAUUGCUUUUGGUGGCCCCAGCUUACAGCUUCAACUGCCUUGGAAUG AGCAACAGAGACUUCUUGGAAGGAGUGUCUGGAGCAACAUGGGUGGA UUUGGUUCUCGAAGGCGACAGCUGCGUGACUAUCAUGUCUAAGGACA AGCCUACCAUCGAUGUGAAGAUGAUGAAUAUGGAGGCGGCCAACCUG GCAGAGGUCCGCAGUUAUUGCUAUUUGGCUACCGUCAGCGAUCUCUC CACCAAAGCUGCGUGCCCGACCAUGGGAGAAGCUCACAAUGACAAAC GUGCUGACCCAGCUUUUGUGUGCAGACAAGGAGUGGUGGACAGGGGC UGGGGCAACGGCUGCGGACUAUUUGGCAAAGGAAGCAUUGACACAUG CGCCAAAUUUGCCUGCUCUACCAAGGCAAUAGGAAGAACCAUCUUGA AAGAGAAUAUCAAGUAUGAAGUGGCCAUUUUUGUCCAUGGGCCAACU ACUGUGGAGUCGCACGGAAACUACUCCACACAGGCUGGAGCCACUCA GGCAGGGAGAUUCAGCAUCACUCCUGCGGCGCCUUCAUACACACUAA AGCUUGGAGAAUAUGGAGAGGUGACAGUGGACUGUGAACCACGGUCA GGGAUUGACACCAAUGCAUACUACGUGAUGACUGUUGGAACAAAGAC GUUCUUGGUCCAUCGUGAGUGGUUCAUGGACCUCAACCUCCCUUGGA GCAGUGCUGGAAGUACUGUGUGGAGGAACAGAGAGACGUUAAUGGAG UUUGAGGAACCGCACGCCACGAAGCAGUCUGUGAUAGCAUUGGGCUC ACAAGAGGGAGCUCUGCAUCAAGCUUUGGCUGGAGCCAUUCCUGUGG AAUUUUCAAGCAACACUGUCAAGUUGACGUCGGGUCAUUUGAAGUGU AGAGUGAAGAUGGAAAAAUUGCAGUUGAAGGGAACAACCUAUGGCGU CUGUUCAAAGGCUUUCAAGUUUCUUGGGACUCCCGCAGACACAGGUC ACGGCACUGUGGUGUUGGAAUUGCAGUACACUGGCACGGACGGACCU UGCAAAGUUCCUAUCUCGUCAGUGGCUUCAUUGAACGACCUAACGCC AGUGGGUAGAUUGGUCACUGUCAACCCUUUUGUUUCAGUGGCCACGG CCAACGCUAAGGUCCUGAUUGAAUUGGAACCACCCUUUGGAGACUCA UACAUAGUGGUGGGCAGAGGAGAACAACAGAUCAAUCAUCAUUGGCA CAAGUCUGGAAGCAGCAUUGGCAAAGCCUUUACAACCACCCUCAAAG GAGCGCAGAGACUAGCCGCUCUAGGAGACACAGCUUGGGACUUUGGA UCAGUUGGAGGGGUGUUCACCUCAGUUGGGAAGGCUGUCCAUCAAGU GUUCGGAGGAGCAUUCCGCUCACUGUUCGGAGGCAUGUCCUGGAUAA CGCAAGGAUUGCUGGGGGCUCUCUUGUUGUGGAUGGGCAUCAAUGCU CGUGAUAGGUCCAUAGCUCUCACGUUUCUCGCAGUUGGAGGAGUUCU GCUCUUCCUCUCCGUGAACGUGCAUGCU SE_WNV_TX8 ORF: 36 559_E2E1 AUGUUCAACUGCCUUGGAAUGAGCAACAGAGACUUCUUGGAAGGAGU WNV, USA GUCUGGAGCAACAUGGGUGGAUUUGGUUCUCGAAGGCGACAGCUGCG isolate UGACUAUCAUGUCUAAGGACAAGCCUACCAUCGAUGUGAAGAUGAUG AAUAUGGAGGCGGCCAACCUGGCAGAGGUCCGCAGUUAUUGCUAUUU GGCUACCGUCAGCGAUCUCUCCACCAAAGCUGCGUGCCCGACCAUGG GAGAAGCUCACAAUGACAAACGUGCUGACCCAGCUUUUGUGUGCAGA CAAGGAGUGGUGGACAGGGGCUGGGGCAACGGCUGCGGACUAUUUGG CAAAGGAAGCAUUGACACAUGCGCCAAAUUUGCCUGCUCUACCAAGG CAAUAGGAAGAACCAUCUUGAAAGAGAAUAUCAAGUAUGAAGUGGCC AUUUUUGUCCAUGGGCCAACUACUGUGGAGUCGCACGGAAACUACUC CACACAGGCUGGAGCCACUCAGGCAGGGAGAUUCAGCAUCACUCCUG CGGCGCCUUCAUACACACUAAAGCUUGGAGAAUAUGGAGAGGUGACA GUGGACUGUGAACCACGGUCAGGGAUUGACACCAAUGCAUACUACGU GAUGACUGUUGGAACAAAGACGUUCUUGGUCCAUCGUGAGUGGUUCA UGGACCUCAACCUCCCUUGGAGCAGUGCUGGAAGUACUGUGUGGAGG AACAGAGAGACGUUAAUGGAGUUUGAGGAACCGCACGCCACGAAGCA GUCUGUGAUAGCAUUGGGCUCACAAGAGGGAGCUCUGCAUCAAGCUU UGGCUGGAGCCAUUCCUGUGGAAUUUUCAAGCAACACUGUCAAGUUG ACGUCGGGUCAUUUGAAGUGUAGAGUGAAGAUGGAAAAAUUGCAGUU GAAGGGAACAACCUAUGGCGUCUGUUCAAAGGCUUUCAAGUUUCUUG GGACUCCCGCAGACACAGGUCACGGCACUGUGGUGUUGGAAUUGCAG UACACUGGCACGGACGGACCUUGCAAAGUUCCUAUCUCGUCAGUGGC UUCAUUGAACGACCUAACGCCAGUGGGUAGAUUGGUCACUGUCAACC CUUUUGUUUCAGUGGCCACGGCCAACGCUAAGGUCCUGAUUGAAUUG GAACCACCCUUUGGAGACUCAUACAUAGUGGUGGGCAGAGGAGAACA ACAGAUCAAUCAUCAUUGGCACAAGUCUGGAAGCAGCAUUGGCAAAG CCUUUACAACCACCCUCAAAGGAGCGCAGAGACUAGCCGCUCUAGGA GACACAGCUUGGGACUUUGGAUCAGUUGGAGGGGUGUUCACCUCAGU UGGGAAGGCUGUCCAUCAAGUGUUCGGAGGAGCAUUCCGCUCACUGU UCGGAGGCAUGUCCUGGAUAACGCAAGGAUUGCUGGGGGCUCUCUUG UUGUGGAUGGGCAUCAAUGCUCGUGAUAGGUCCAUAGCUCUCACGUU UCUCGCAGUUGGAGGAGUUCUGCUCUUCCUCUCCGUGAACGUGCAUG CU SE_WNV_TX8 mRNA sequence: 37 559_E2E1 G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCA WNV, USA CCAUGUUCAACUGCCUUGGAAUGAGCAACAGAGACUUCUUGGAAGGA isolate GUGUCUGGAGCAACAUGGGUGGAUUUGGUUCUCGAAGGCGACAGCUG CGUGACUAUCAUGUCUAAGGACAAGCCUACCAUCGAUGUGAAGAUGA UGAAUAUGGAGGCGGCCAACCUGGCAGAGGUCCGCAGUUAUUGCUAU UUGGCUACCGUCAGCGAUCUCUCCACCAAAGCUGCGUGCCCGACCAU GGGAGAAGCUCACAAUGACAAACGUGCUGACCCAGCUUUUGUGUGCA GACAAGGAGUGGUGGACAGGGGCUGGGGCAACGGCUGCGGACUAUUU GGCAAAGGAAGCAUUGACACAUGCGCCAAAUUUGCCUGCUCUACCAA GGCAAUAGGAAGAACCAUCUUGAAAGAGAAUAUCAAGUAUGAAGUGG CCAUUUUUGUCCAUGGGCCAACUACUGUGGAGUCGCACGGAAACUAC UCCACACAGGCUGGAGCCACUCAGGCAGGGAGAUUCAGCAUCACUCC UGCGGCGCCUUCAUACACACUAAAGCUUGGAGAAUAUGGAGAGGUGA CAGUGGACUGUGAACCACGGUCAGGGAUUGACACCAAUGCAUACUAC GUGAUGACUGUUGGAACAAAGACGUUCUUGGUCCAUCGUGAGUGGUU CAUGGACCUCAACCUCCCUUGGAGCAGUGCUGGAAGUACUGUGUGGA GGAACAGAGAGACGUUAAUGGAGUUUGAGGAACCGCACGCCACGAAG CAGUCUGUGAUAGCAUUGGGCUCACAAGAGGGAGCUCUGCAUCAAGC UUUGGCUGGAGCCAUUCCUGUGGAAUUUUCAAGCAACACUGUCAAGU UGACGUCGGGUCAUUUGAAGUGUAGAGUGAAGAUGGAAAAAUUGCAG UUGAAGGGAACAACCUAUGGCGUCUGUUCAAAGGCUUUCAAGUUUCU UGGGACUCCCGCAGACACAGGUCACGGCACUGUGGUGUUGGAAUUGC AGUACACUGGCACGGACGGACCUUGCAAAGUUCCUAUCUCGUCAGUG GCUUCAUUGAACGACCUAACGCCAGUGGGUAGAUUGGUCACUGUCAA CCCUUUUGUUUCAGUGGCCACGGCCAACGCUAAGGUCCUGAUUGAAU UGGAACCACCCUUUGGAGACUCAUACAUAGUGGUGGGCAGAGGAGAA CAACAGAUCAAUCAUCAUUGGCACAAGUCUGGAAGCAGCAUUGGCAA AGCCUUUACAACCACCCUCAAAGGAGCGCAGAGACUAGCCGCUCUAG GAGACACAGCUUGGGACUUUGGAUCAGUUGGAGGGGUGUUCACCUCA GUUGGGAAGGCUGUCCAUCAAGUGUUCGGAGGAGCAUUCCGCUCACU GUUCGGAGGCAUGUCCUGGAUAACGCAAGGAUUGCUGGGGGCUCUCU UGUUGUGGAUGGGCAUCAAUGCUCGUGAUAGGUCCAUAGCUCUCACG UUUCUCGCAGUUGGAGGAGUUCUGCUCUUCCUCUCCGUGAACGUGCA UGCUUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUU GGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCGU GGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG SE_WNV_TX8 ORF: 38 559_E2E1_SP AUGCAGAGAGUUGUGUUUGUCGUGCUAUUGCUUUUGGUGGCCCCAGC WNV, USA UUACAGCUUCAACUGCCUUGGAAUGAGCAACAGAGACUUCUUGGAAG isolate GAGUGUCUGGAGCAACAUGGGUGGAUUUGGUUCUCGAAGGCGACAGC UGCGUGACUAUCAUGUCUAAGGACAAGCCUACCAUCGAUGUGAAGAU GAUGAAUAUGGAGGCGGCCAACCUGGCAGAGGUCCGCAGUUAUUGCU AUUUGGCUACCGUCAGCGAUCUCUCCACCAAAGCUGCGUGCCCGACC AUGGGAGAAGCUCACAAUGACAAACGUGCUGACCCAGCUUUUGUGUG CAGACAAGGAGUGGUGGACAGGGGCUGGGGCAACGGCUGCGGACUAU UUGGCAAAGGAAGCAUUGACACAUGCGCCAAAUUUGCCUGCUCUACC AAGGCAAUAGGAAGAACCAUCUUGAAAGAGAAUAUCAAGUAUGAAGU GGCCAUUUUUGUCCAUGGGCCAACUACUGUGGAGUCGCACGGAAACU ACUCCACACAGGCUGGAGCCACUCAGGCAGGGAGAUUCAGCAUCACU CCUGCGGCGCCUUCAUACACACUAAAGCUUGGAGAAUAUGGAGAGGU GACAGUGGACUGUGAACCACGGUCAGGGAUUGACACCAAUGCAUACU ACGUGAUGACUGUUGGAACAAAGACGUUCUUGGUCCAUCGUGAGUGG UUCAUGGACCUCAACCUCCCUUGGAGCAGUGCUGGAAGUACUGUGUG GAGGAACAGAGAGACGUUAAUGGAGUUUGAGGAACCGCACGCCACGA AGCAGUCUGUGAUAGCAUUGGGCUCACAAGAGGGAGCUCUGCAUCAA GCUUUGGCUGGAGCCAUUCCUGUGGAAUUUUCAAGCAACACUGUCAA GUUGACGUCGGGUCAUUUGAAGUGUAGAGUGAAGAUGGAAAAAUUGC AGUUGAAGGGAACAACCUAUGGCGUCUGUUCAAAGGCUUUCAAGUUU CUUGGGACUCCCGCAGACACAGGUCACGGCACUGUGGUGUUGGAAUU GCAGUACACUGGCACGGACGGACCUUGCAAAGUUCCUAUCUCGUCAG UGGCUUCAUUGAACGACCUAACGCCAGUGGGUAGAUUGGUCACUGUC AACCCUUUUGUUUCAGUGGCCACGGCCAACGCUAAGGUCCUGAUUGA AUUGGAACCACCCUUUGGAGACUCAUACAUAGUGGUGGGCAGAGGAG AACAACAGAUCAAUCAUCAUUGGCACAAGUCUGGAAGCAGCAUUGGC AAAGCCUUUACAACCACCCUCAAAGGAGCGCAGAGACUAGCCGCUCU AGGAGACACAGCUUGGGACUUUGGAUCAGUUGGAGGGGUGUUCACCU CAGUUGGGAAGGCUGUCCAUCAAGUGUUCGGAGGAGCAUUCCGCUCA CUGUUCGGAGGCAUGUCCUGGAUAACGCAAGGAUUGCUGGGGGCUCU CUUGUUGUGGAUGGGCAUCAAUGCUCGUGAUAGGUCCAUAGCUCUCA CGUUUCUCGCAGUUGGAGGAGUUCUGCUCUUCCUCUCCGUGAACGUG CAUGCU SE_WNV_TX8 mRNA sequence: 39 559_E2E1_SP G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCA WNV, USA CCAUGCAGAGAGUUGUGUUUGUCGUGCUAUUGCUUUUGGUGGCCCCA isolate GCUUACAGCUUCAACUGCCUUGGAAUGAGCAACAGAGACUUCUUGGA AGGAGUGUCUGGAGCAACAUGGGUGGAUUUGGUUCUCGAAGGCGACA GCUGCGUGACUAUCAUGUCUAAGGACAAGCCUACCAUCGAUGUGAAG AUGAUGAAUAUGGAGGCGGCCAACCUGGCAGAGGUCCGCAGUUAUUG CUAUUUGGCUACCGUCAGCGAUCUCUCCACCAAAGCUGCGUGCCCGA CCAUGGGAGAAGCUCACAAUGACAAACGUGCUGACCCAGCUUUUGUG UGCAGACAAGGAGUGGUGGACAGGGGCUGGGGCAACGGCUGCGGACU AUUUGGCAAAGGAAGCAUUGACACAUGCGCCAAAUUUGCCUGCUCUA CCAAGGCAAUAGGAAGAACCAUCUUGAAAGAGAAUAUCAAGUAUGAA GUGGCCAUUUUUGUCCAUGGGCCAACUACUGUGGAGUCGCACGGAAA CUACUCCACACAGGCUGGAGCCACUCAGGCAGGGAGAUUCAGCAUCA CUCCUGCGGCGCCUUCAUACACACUAAAGCUUGGAGAAUAUGGAGAG GUGACAGUGGACUGUGAACCACGGUCAGGGAUUGACACCAAUGCAUA CUACGUGAUGACUGUUGGAACAAAGACGUUCUUGGUCCAUCGUGAGU GGUUCAUGGACCUCAACCUCCCUUGGAGCAGUGCUGGAAGUACUGUG UGGAGGAACAGAGAGACGUUAAUGGAGUUUGAGGAACCGCACGCCAC GAAGCAGUCUGUGAUAGCAUUGGGCUCACAAGAGGGAGCUCUGCAUC AAGCUUUGGCUGGAGCCAUUCCUGUGGAAUUUUCAAGCAACACUGUC AAGUUGACGUCGGGUCAUUUGAAGUGUAGAGUGAAGAUGGAAAAAUU GCAGUUGAAGGGAACAACCUAUGGCGUCUGUUCAAAGGCUUUCAAGU UUCUUGGGACUCCCGCAGACACAGGUCACGGCACUGUGGUGUUGGAA UUGCAGUACACUGGCACGGACGGACCUUGCAAAGUUCCUAUCUCGUC AGUGGCUUCAUUGAACGACCUAACGCCAGUGGGUAGAUUGGUCACUG UCAACCCUUUUGUUUCAGUGGCCACGGCCAACGCUAAGGUCCUGAUU GAAUUGGAACCACCCUUUGGAGACUCAUACAUAGUGGUGGGCAGAGG AGAACAACAGAUCAAUCAUCAUUGGCACAAGUCUGGAAGCAGCAUUG GCAAAGCCUUUACAACCACCCUCAAAGGAGCGCAGAGACUAGCCGCU CUAGGAGACACAGCUUGGGACUUUGGAUCAGUUGGAGGGGUGUUCAC CUCAGUUGGGAAGGCUGUCCAUCAAGUGUUCGGAGGAGCAUUCCGCU CACUGUUCGGAGGCAUGUCCUGGAUAACGCAAGGAUUGCUGGGGGCU CUCUUGUUGUGGAUGGGCAUCAAUGCUCGUGAUAGGUCCAUAGCUCU CACGUUUCUCGCAGUUGGAGGAGUUCUGCUCUUCCUCUCCGUGAACG UGCAUGCUUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCC CCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCC CCGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG

For all sequences in Table 8:

5′UTR: DNA (SEQ ID NO: 40) TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAA ATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACC 3′UTR: DNA (SEQ ID NO: 41) TGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTC CCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCGTGGTCTTTGAA TAAAGTCTGAGTGGGCGGC 5′UTR: mRNA (SEQ ID NO: 42) UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAA AUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACC 3′UTR: mRNA (SEQ ID NO: 43) UGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUC CCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCGUGGUCUUUGAA UAAAGUCUGAGUGGGCGGC

TABLE10 WNVAmino Acid Sequences Antigen Amino Acid Sequence SEQ ID NO: SE_WNV_TX8 MFNCLGMSNRDFLEGVSGATWVDLVLEGDSCVTIMSKDKPTIDVKM 44 559_E2E1 MNMEAANLAEVRSYCYLATVSDLSTKAACPTMGEAHNDKRADPAFV CRQGVVDRGWGNGCGLFGKGSIDTCAKFACSTKAIGRTILKENIKY EVAIFVHGPTTVESHGNYSTQAGATQAGRFSITPAAPSYTLKLGEY GEVTVDCEPRSGIDTNAYYVMTVGTKTFLVHREWFMDLNLPWSSAG STVWRNRETLMEFEEPHATKQSINVIALGSQEGALHQALAGAIPVE FSSNTVKLTSGHLKCRVKMEKLQLKGTTYGVCSKAFKFLGTPADTG HGTVVLELQYTGTDGPCKVPISSINVASLNDLTPVGRLVTVNPFVS INVATANAKVLIELEPPFGDSYIVVGRGEQQINHHWHKSGSSIGKA FTTTLKGAQRLAALGDTAWDFGSINVGGVFTSINVGKAVHQVFGGA FRSLFGGMSWITQGLLGALLLWMGINARDRSIALTFLAVGGVLLFL SINVNVHA SE_WNV_TX8 MQRVVFVVLLLLVAPAYSFNCLGMSNRDFLEGVSGATWVDLVLEGD 45 559_E2E1_SP SCVTIMSKDKPTIDVKMMNMEAANLAEVRSYCYLATVSDLSTKAAC PTMGEAHNDKRADPAFVCRQGVVDRGWGNGCGLFGKGSIDTCAKFA CSTKAIGRTILKENIKYEVAIFVHGPTTVESHGNYSTQAGATQAGR FSITPAAPSYTLKLGEYGEVTVDCEPRSGIDTNAYYVMTVGTKTFL VHREWFMDLNLPWSSAGSTVWRNRETLMEFEEPHATKQSINVIALG SQEGALHQALAGAIPVEFSSNTVKLTSGHLKCRVKMEKLQLKGTTY GVCSKAFKFLGTPADTGHGTVVLELQYTGTDGPCKVPISSINVASL NDLTPVGRLVTVNPFVSINVATANAKVLIELEPPFGDSYIVVGRGE QQINHHWHKSGSSIGKAFTTTLKGAQRLAALGDTAWDFGSINVGGV FTSINVGKAVHQVFGGAFRSLFGGMSWITQGLLGALLLWMGINARD RSIALTFLAVGGVLLFLSINVNVHA SE_West Nile MWLVSLAIVTACAGAMKLSNFQGKVMMTVNATDVTDVITIPTAAGK 46 PRM-E NLCIVRAMDVGYMCDDTITYECPVLSAGNDPEDIDCWCTKSAVYVR YGRCTKTRHSRRSRRSLTVQTHGESTLANKKGAWMDSTKATRYLVK TESWILRNPGYALVAAVIGWMLGSNTMQRVVFVVLLLLVAPAYSFN CLGMSNRDFLEGVSGATWVDLVLEGDSCVTIMSKDKPTIDVKMMNM EAANLAEVRSYCYLATVSDLSTKAACPTMGEAHNDKRADPAFVCRQ GVVDRGWGNGCGLFGKGSIDTCAKFACSTKAIGRTILKENIKYEVA IFVHGPTTVESHGNYSTQAGATQAGRFSITPAAPSYTLKLGEYGEV TVDCEPRSGIDTNAYYVMTVGTKTFLVHREWFMDLNLPWSSAGSTV WRNRETLMEFEEPHATKQSINVIALGSQEGALHQALAGAIPVEFSS NTVKLTSGHLKCRVKMEKLQLKGTTYGVCSKAFKFLGTPADTGHGT VVLELQYTGTDGPCKVPISSINVASLNDLTPVGRLVTVNPFVSINV ATANAKVLIELEPPFGDSYIVVGRGEQQINHHWHKSGSSIGKAFTT TLKGAQRLAALGDTAWDFGSINVGGVFTSINVGKAVHQVFGGAFRS LFGGMSWITQGLLGALLLWMGINARDRSIALTFLAVGGVLLFLSIN VNVHA SE_WNV_TX8 MFNCLGMSNRDFLEGVSGATWVDLVLEGDSCVTIMSKDKPTIDVKM 47 559_E2E1 MNMEAANLAEVRSYCYLATVSDLSTKAACPTMGEAHNDKRADPAFV CRQGVVDRGWGNGCGLFGKGSIDTCAKFACSTKAIGRTILKENIKY EVAIFVHGPTTVESHGNYSTQAGATQAGRFSITPAAPSYTLKLGEY GEVTVDCEPRSGIDTNAYYVMTVGTKTFLVHREWFMDLNLPWSSAG STVWRNRETLMEFEEPHATKQSINVIALGSQEGALHQALAGAIPVE FSSNTVKLTSGHLKCRVKMEKLQLKGTTYGVCSKAFKFLGTPADTG HGTVVLELQYTGTDGPCKVPISSINVASLNDLTPVGRLVTVNPFVS INVATANAKVLIELEPPFGDSYIVVGRGEQQINHHWHKSGSSIGKA FTTTLKGAQRLAALGDTAWDFGSINVGGVFTSINVGKAVHQVFGGA FRSLFGGMSWITQGLLGALLLWMGINARDRSIALTFLAVGGVLLFL SINVNVHA Underlined sequence corresponds to a signal peptide, which may be omitted from each sequence. Thus, any RNAvaccine provided herein may encode an antigen represented by a sequence of Table 10, with or without the underlined signal peptide.

TABLE 11 WNV strains/isolates, Envelope proteins/variants - Homo sapiens GenBank Collection Accession Length Type Country Genome regions date Virus name ABR19639 3434 2 South UTR5CprMENS1NS2 1958 West Nile virus H442, Africa ANS2BNS3NS4A2KN complete genome S4BNS5 ABR19638 3434 2 South UTR5CprMENS1NS2 2000 West Nile virus Africa ANS2BNS3NS4A2KN SA381/00, complete S4BNS5 genome ABR19637 3434 2 South UTR5CprMENS1NS2 2001 West Nile virus Africa ANS2BNS3NS4A2KN SA93/01, complete S4BNS5 genome ABR19636 3434 2 South UTR5CprMENS1NS2 1989 West Nile virus Africa ANS2BNS3NS4A2KN SPU116/89, complete S4BNS5 genome AAK06624 3433 1A Russia UTR5CprMENS1NS2 West Nile virus VLG-4 ANS2BNS3NS4A2KN polyprotein precursor, S4BNS5UTR3 gene, complete cds AAV54504 3433 1A USA UTR5CprMENS1NS2 2002 West Nile virus from ANS2BNS3NS4A2KN USA, complete genome S4BNS5 ACV90471 3433 1A USA CprMENS1NS2ANS2 Aug. 3, 2005 West Nile virus isolate BNS3NS4A2KNS4BN 007WG-TX05EP S5 polyprotein gene, complete cds ACV90472 3433 1A USA CprMENS1NS2ANS2 Sep. 1, 2005 West Nile virus isolate BNS3NS4A2KNS4BN 009WG-NM05LC S5 polyprotein gene, complete cds ACV90473 3433 1A USA CprMENS1NS2ANS2 Aug. 30, 2006 West Nile virus isolate BNS3NS4A2KNS4BN 011WG-TX06EP S5 polyprotein gene, complete cds ACV90474 3433 1A USA CprMENS1NS2ANS2 Jun. 23, 2007 West Nile virus isolate BNS3NS4A2KNS4BN 013WG-TX07EP S5UTR3 polyprotein gene, complete cds ACV90475 3433 1A USA CprMENS1NS2ANS2 Jul. 14, 2003 West Nile virus isolate BNS3NS4A2KNS4BN 024WG-CA03OR S5UTR3 polyprotein gene, complete cds ABD85067 3433 1A USA CprMENS1NS2ANS2 2003 West Nile virus isolate BNS3NS4A2KNS4BN 03-104WI polyprotein S5 precursor, gene, complete cds ABD85068 3433 1A USA CprMENS1NS2ANS2 2003 West Nile virus isolate BNS3NS4A2KNS4BN 03-113FL polyprotein S5 precursor, gene, complete cds ABD85069 3433 1A USA CprMENS1NS2ANS2 2003 West Nile virus isolate BNS3NS4A2KNS4BN 03-120FL polyprotein S5 precursor, gene, complete cds ABD85070 3433 1A USA CprMENS1NS2ANS2 2003 West Nile virus isolate BNS3NS4A2KNS4BN 03-124FL polyprotein S5 precursor, gene, complete cds ABD85064 3433 1A USA CprMENS1NS2ANS2 2003 West Nile virus isolate BNS3NS4A2KNS4BN 03-20TX polyprotein S5 precursor, gene, complete cds ABD85065 3433 1A USA CprMENS1NS2ANS2 2003 West Nile virus isolate BNS3NS4A2KNS4BN 03-22TX polyprotein S5 precursor, gene, complete cds ABD85066 3433 1A USA CprMENS1NS2ANS2 2003 West Nile virus isolate BNS3NS4A2KNS4BN 03-82IL polyprotein S5 precursor, gene, complete cds ABD85071 3433 1A USA CprMENS1NS2ANS2 2004 West Nile virus isolate BNS3NS4A2KNS4BN 04-213CA polyprotein S5 precursor, gene, complete cds ABD85072 3433 1A USA CprMENS1NS2ANS2 2004 West Nile virus isolate BNS3NS4A2KNS4BN 04-214CO polyprotein S5 precursor, gene, complete cds ABD85073 3433 1A USA CprMENS1NS2ANS2 2004 West Nile virus isolate BNS3NS4A2KNS4BN 04-216CO polyprotein S5 precursor, gene, complete cds ABD85074 3433 1A USA CprMENS1NS2ANS2 2004 West Nile virus isolate BNS3NS4A2KNS4BN 04-218CO polyprotein S5 precursor, gene, complete cds ABD85075 3433 1A USA CprMENS1NS2ANS2 2004 West Nile virus isolate BNS3NS4A2KNS4BN 04-219CO polyprotein S5 precursor, gene, complete cds ABD85076 3433 1A USA CprMENS1NS2ANS2 2004 West Nile virus isolate BNS3NS4A2KNS4BN 04-233ND polyprotein S5UTR3 precursor, gene, complete cds ABD85077 3433 1A USA CprMENS1NS2ANS2 2004 West Nile virus isolate BNS3NS4A2KNS4BN 04-236NM polyprotein S5UTR3 precursor, gene, complete cds ABD85078 3433 1A USA CprMENS1NS2ANS2 2004 West Nile virus isolate BNS3NS4A2KNS4BN 04-237NM polyprotein S5 precursor, gene, complete cds ABD85079 3433 1A USA CprMENS1NS2ANS2 2004 West Nile virus isolate BNS3NS4A2KNS4BN 04-238CA polyprotein S5 precursor, gene, complete cds ABD85080 3433 1A USA CprMENS1NS2ANS2 2004 West Nile virus isolate BNS3NS4A2KNS4BN 04-240CA polyprotein S5 precursor, gene, complete cds ABD85081 3433 1A USA CprMENS1NS2ANS2 2004 West Nile virus isolate BNS3NS4A2KNS4BN 04-244CA polyprotein S5UTR3 precursor, gene, complete cds ABD85082 3433 1A USA CprMENS1NS2ANS2 2004 West Nile virus isolate BNS3NS4A2KNS4BN 04-251AZ polyprotein S5 precursor, gene, complete cds ABD85083 3433 1A USA CprMENS1NS2ANS2 2004 West Nile virus isolate BNS3NS4A2KNS4BN 04-252AZ polyprotein S5 precursor, gene, complete cds ACV90476 3433 1A USA CprMENS1NS2ANS2 Jul. 24, 2004 West Nile virus isolate BNS3NS4A2KNS4BN 080WG-CA04LA S5UTR3 polyprotein gene, complete cds ACV90477 3433 1A USA CprMENS1NS2ANS2 Aug. 27, 2004 West Nile virus isolate BNS3NS4A2KNS4BN 091WG-CA04SB S5UTR3 polyprotein gene, complete cds ACV90478 3433 1A USA CprMENS1NS2ANS2 Jun. 27, 2005 West Nile virus isolate BNS3NS4A2KNS4BN 099WG-CA05SB S5UTR3 polyprotein gene, complete cds ACV90479 3433 1A USA CprMENS1NS2ANS2 Jul. 27, 2005 West Nile virus isolate BNS3NS4A2KNS4BN 101WG-CA05SB S5UTR3 polyprotein gene, complete cds ACV90480 3433 1A USA CprMENS1NS2ANS2 Aug. 1, 2005 West Nile virus isolate BNS3NS4A2KNS4BN 103WG-CA05LA S5UTR3 polyprotein gene, complete cds AGI16461 3433 1A India CprMENS1NS2ANS2 2011/02 West Nile virus isolate BNS3NS4A2KNS4BN 1048813, complete S5UTR3 genome ACV90481 3433 1A USA CprMENS1NS2ANS2 Aug. 26, 2005 West Nile virus isolate BNS3NS4A2KNS4BN 116WG-CA05LA S5UTR3 polyprotein gene, complete cds ACV90482 3433 1A USA CprMENS1NS2ANS2 Sep. 9, 2005 West Nile virus isolate BNS3NS4A2KNS4BN 124WG-AZ05PI S5UTR3 polyprotein gene, complete cds ACV90483 3433 1A USA CprMENS1NS2ANS2 Oct. 4, 2005 West Nile virus isolate BNS3NS4A2KNS4BN 132WG-CA05LA S5UTR3 polyprotein gene, complete cds ACV90484 3433 1A USA CprMENS1NS2ANS2 Aug. 22, 2006 West Nile virus isolate BNS3NS4A2KNS4BN 142WG-NE06DO S5UTR3 polyprotein gene, complete cds ACV90485 3433 1A USA CprMENS1NS2ANS2 Sep. 3, 2006 West Nile virus isolate BNS3NS4A2KNS4BN 144WG-AZ06PI S5UTR3 polyprotein gene, complete cds ACV90486 3433 1A USA CprMENS1NS2ANS2 Aug. 2, 2007 West Nile virus isolate BNS3NS4A2KNS4BN 148WG-CA07LA S5UTR3 polyprotein gene, complete cds ACV90487 3433 1A USA CprMENS1NS2ANS2 Aug. 16, 2007 West Nile virus isolate BNS3NS4A2KNS4BN 149WG-CA07LA S5UTR3 polyprotein gene, complete cds ABA54590 3433 1A USA UTR5CprMENS1NS2 2004 West Nile virus isolate ANS2BNS3NS4A2KN AZ 2004, complete S4BNS5 genome AAP22086 3433 1A Russia UTR5CprMENS1NS2 1999 West Nile virus isolate ANS2BNS3NS4A2KN Ast99-901, complete S4BNS5UTR3 genome ABG81343 3433 1A USA UTR5CprMENS1NS2 2005 West Nile virus isolate ANS2BNS3NS4A2KN BSL13-2005, complete S4BNS5 genome ABG81344 3433 1A USA UTR5CprMENS1NS2 2005 West Nile virus isolate ANS2BNS3NS4A2KN BSL2-2005, complete S4BNS5UTR3 genome AFJ05105 3433 1A USA UTR5CprMENS1NS2 2011 West Nile virus isolate ANS2BNS3NS4A2KN BSL23-11, complete S4BNS5 genome AFJ05106 3433 1A USA UTR5CprMENS1NS2 2011 West Nile virus isolate ANS2BNS3NS4A2KN BSL24-11, complete S4BNS5 genome AFJ05107 3433 1A USA UTR5CprMENS1NS2 2011 West Nile virus isolate ANS2BNS3NS4A2KN BSL26-11, complete S4BNS5 genome AFJ05103 3433 1A USA UTR5CprMENS1NS2 2011 West Nile virus isolate ANS2BNS3NS4A2KN BSL4-11, complete S4BNS5UTR3 genome ABG81340 3433 1A USA UTR5CprMENS1NS2 2004 West Nile virus isolate ANS2BNS3NS4A2KN BSL5-2004, complete S4BNS5 genome AFJ05104 3433 1A USA UTR5CprMENS1NS2 2011 West Nile virus isolate ANS2BNS3NS4A2KN BSL6-11, complete S4BNS5 genome AAY55949 3433 1A USA UTR5CprMENS1NS2 2003 West Nile virus isolate ANS2BNS3NS4A2KN FDA-BSL5-2003, S4BNS5 complete genome ABA54585 3433 1A USA UTR5CprMENS1NS2 2002 West Nile virus isolate ANS2BNS3NS4A2KN GA 2002 1, complete S4BNS5 genome ABA54586 3433 1A USA UTR5CprMENS1NS2 2002 West Nile virus isolate ANS2BNS3NS4A2KN GA 2002 2, complete S4BNS5 genome ABG81341 3433 1A USA UTR5CprMENS1NS2 2005 West Nile virus isolate ANS2BNS3NS4A2KN GCTX1-2005, complete S4BNS5 genome ABG81342 3433 1A USA UTR5CprMENS1NS2 2005 West Nile virus isolate ANS2BNS3NS4A2KN GCTX2-2005, complete S4BNS5 genome ABA54589 3433 1A USA UTR5CprMENS1NS2 2002 West Nile virus isolate ANS2BNS3NS4A2KN IN 2002, complete S4BNS5 genome AEV45199 3433 1A Italy UTR5CprMENS1NS2 2011/09 West Nile virus isolate ANS2BNS3NS4A2KN Italy/2011/AN-1, S4BNS5UTR3 complete genome AEV45200 3434 2 Italy UTR5CprMENS1NS2 2011/09 West Nile virus isolate ANS2BNS3NS4A2KN Italy/2011/AN-2, S4BNS5 complete genome AAP22087 3433 1A Russia UTR5CprMENS1NS2 2000 West Nile virus isolate ANS2BNS3NS4A2KN LEIV-Vlg00-27924, S4BNS5UTR3 complete genome AAP22089 3433 1A Russia UTR5CprMENS1NS2 1999 West Nile virus isolate ANS2BNS3NS4A2KN LEIV-Vlg99-27889, S4BNS5UTR3 complete genome AIY22515 3433 1A USA UTR5CprMENS1NS2 2001 West Nile virus isolate ANS2BNS3NS4A2KN NY2001-6263, complete S4BNS5 genome ABA54591 3433 1A USA UTR5CprMENS1NS2 2002 West Nile virus isolate ANS2BNS3NS4A2KN OH 2002, complete S4BNS5 genome ACJ67802 3434 2 Russia UTR5CprMENS1NS2 2007 West Nile virus isolate ANS2BNS3NS4A2KN Reb_VLG_07_H S4BNS5UTR3 polyprotein gene, complete cds ABU41789 3434 2 South CprMENS1NS2ANS2 1989 West Nile virus isolate Africa BNS3NS4A2KNS4BN SPU116-89 polyprotein S5 gene, complete cds ABA54587 3433 1A USA UTR5CprMENS1NS2 2002 West Nile virus isolate ANS2BNS3NS4A2KN TX 2002 1, complete S4BNS5 genome ABA54594 3433 1A USA UTR5CprMENS1NS2 2002 West Nile virus isolate ANS2BNS3NS4A2KN TX 2002 2, complete S4BNS5 genome ABA54588 3433 1A USA UTR5CprMENS1NS2 2003 West Nile virus isolate ANS2BNS3NS4A2KN TX 2003, complete S4BNS5 genome ADI33159 3433 1A Israel UTR5CprMENS1NS2 2000 West Nile virus isolate ANS2BNS3NS4A2KN WNV_0043h_ISR00 S4BNS5UTR3 from Israel polyprotein gene, complete cds ADI33161 3433 1A Israel UTR5CprMENS1NS2 2000 West Nile virus isolate ANS2BNS3NS4A2KN WNV_0304h_ISR00 S4BNS5UTR3 from Israel polyprotein gene, complete cds ADN97107 3433 1A Israel UTR5CprMENS1NS2 1953 West Nile virus isolate ANS2BNS3NS4A2KN twn9, complete genome S4BNS5 AAN85090 3433 1A USA UTR5CprMENS1NS2 2001 West Nile virus ANS2BNS3NS4A2KN polyprotein precursor, S4BNS5 gene, complete cds ABC40712 3433 5 India UTR5CprMENS1NS2 1980 West Nile virus strain ANS2BNS3NS4A2KN 804994, complete S4BNS5UTR3 genome AFO64356 3433 1A Russia UTR5CprMENS1NS2 1999 West Nile virus strain ANS2BNS3NS4A2KN Ast-986, complete S4BNS5UTR3 genome AIW82672 3434 2 Austria UTR5CprMENS1NS2 2014 West Nile virus strain ANS2BNS3NS4A2KN BD-AUT, complete S4BNS5UTR3 genome AGX89730 3434 2 Greece UTR5CprMENS1NS2 2012/09 West Nile virus strain ANS2BNS3NS4A2KN Greece/2012/Kavala/39.1, S4BNS5UTR3 complete genome AIA23852 3434 2 Greece UTR5CprMENS1NS2 2013 West Nile virus strain ANS2BNS3NS4A2KN Greece/2013/C105 S4BNS5UTR3 polyprotein gene, complete cds AIA23853 3434 2 Greece UTR5CprMENS1NS2 2013 West Nile virus strain ANS2BNS3NS4A2KN Greece/2013/C147 S4BNS5UTR3 polyprotein gene, complete cds AJR27175 3434 2 Greece UTR5CprMENS1NS2 2013 West Nile virus strain ANS2BNS3NS4A2KN Greece/2013/Kavala_1, S4BNS5UTR3 genome AJR27181 3434 2 Greece UTR5CprMENS1NS2 2013 West Nile virus strain ANS2BNS3NS4A2KN Greece/2013/Kavala_2, S4BNS5UTR3 complete genome AJR27183 3434 2 Greece UTR5CprMENS1NS2 2013 West Nile virus strain ANS2BNS3NS4A2KN Greece/2013/Thessaloniki_3, S4BNS5UTR3 genome AJR27179 3434 2 Greece UTR5CprMENS1NS2 2013 West Nile virus strain ANS2BNS3NS4A2KN Greece/2013/Thessaloniki_4, S4BNS5UTR3 complete genome AJR27176 3434 2 Greece UTR5CprMENS1NS2 2013 West Nile virus strain ANS2BNS3NS4A2KN Greece/2013/Xanthi_1, S4BNS5UTR3 complete genome AJR27177 3434 2 Greece UTR5CprMENS1NS2 2013 West Nile virus strain ANS2BNS3NS4A2KN Greece/2013/Xanthi_2, S4BNS5UTR3 complete genome AJR27178 3434 2 Greece UTR5CprMENS1NS2 2013 West Nile virus strain ANS2BNS3NS4A2KN Greece/2013/Xanthi_3, S4BNS5UTR3 complete genome AJR27182 3434 2 Greece UTR5CprMENS1NS2 2013 West Nile virus strain ANS2BNS3NS4A2KN Greece/2013/Xanthi_5, S4BNS5UTR3 complete genome AAF18443 3433 1A USA UTR5CprMENS1NS2 1999 West Nile virus strain ANS2BNS3NS4A2KN HNY1999 polyprotein S4BNS5UTR3 (C, prM, E, NS1, NS2a, NS2b, NS3, NS4a, NS4b, NS5) gene, complete cds ACX71000 3433 1A Italy UTR5CprMENS1NS2 2009 West Nile virus strain ANS2BNS3NS4A2KN Ita09, complete genome S4BNS5 AGW24516 3433 1A Italy UTR5CprMENS1NS2 2009 West Nile virus strain ANS2BNS3NS4A2KN Italy/2009/FIN, S4BNS5 complete genome AFP50425 3433 1A Italy UTR5CprMENS1NS2 2011/09 West Nile virus strain ANS2BNS3NS4A2KN Italy/2011/Livenza, S4BNS5UTR3 complete genome AFP50426 3433 1A Italy UTR5CprMENS1NS2 2011/09 West Nile virus strain ANS2BNS3NS4A2KN Italy/2011/Piave, S4BNS5 complete genome AFR36922 3433 1A Italy CprMENS1NS2ANS2 Aug. 3, 2012 West Nile virus strain BNS3NS4A2KNS4BN Italy/2012/Livenza/31.1, S5UTR3 complete genome AGS78392 3433 1A Italy CprMENS1NS2ANS2 2012/09 West Nile virus strain BNS3NS4A2KNS4BN Italy/2012/Livenza/37.1, S5UTR3 complete genome AHB37633 3433 1A Italy CprMENS1NS2ANS2 2013/08 West Nile virus strain BNS3NS4A2KNS4BN Italy/2013/Livenza/35.1, S5 complete genome AJR27899 3434 2 Italy CprMENS1NS2ANS2 2013 West Nile virus strain BNS3NS4A2KNS4BN Italy/2013/Mantova/36.1, S5UTR3 complete genome AHH30722 3434 2 Italy UTR5CprMENS1NS2 2013/10 West Nile virus strain ANS2BNS3NS4A2KN Italy/2013/Mantova/40.1 S4BNS5UTR3 polyprotein gene, complete cds AHB37631 3434 2 Italy UTR5CprMENS1NS2 2013/08 West Nile virus strain ANS2BNS3NS4A2KN Italy/2013/Padova/34.1, S4BNS5UTR3 complete genome AGY34434 3434 2 Italy UTR5CprMENS1NS2 2013/08 West Nile virus strain ANS2BNS3NS4A2KN Italy/2013/Rovigo/32.1, S4BNS5UTR3 complete genome AHB37630 3434 2 Italy UTR5CprMENS1NS2 2013/08 West Nile virus strain ANS2BNS3NS4A2KN Italy/2013/Rovigo/33.1, S4BNS5UTR3 complete genome AHB37629 3434 2 Italy UTR5CprMENS1NS2 2013/08 West Nile virus strain ANS2BNS3NS4A2KN Italy/2013/Rovigo/33.2, S4BNS5UTR3 complete genome AHB37632 3434 2 Italy UTR5CprMENS1NS2 2013/08 West Nile virus strain ANS2BNS3NS4A2KN Italy/2013/Rovigo/35.1, S4BNS5UTR3 complete genome AJR27893 3434 2 Italy UTR5CprMENS1NS2 2014 West Nile virus strain ANS2BNS3NS4A2KN Italy/2014/Cremona2, S4BNS5UTR3 complete genome AJR27896 3434 2 Italy UTR5CprMENS1NS2 2014 West Nile virus strain ANS2BNS3NS4A2KN Italy/2014/Cremona4, S4BNS5UTR3 complete genome AJR27892 3434 2 Italy UTR5CprMENS1NS2 2014 West Nile virus strain ANS2BNS3NS4A2KN Italy/2014/Pavia1, S4BNS5UTR3 complete genome AJR27898 3434 2 Italy CprMENS1NS2ANS2 2014 West Nile virus strain BNS3NS4A2KNS4BN Italy/2014/Pavia4, S5UTR3 complete genome AJR27897 3434 2 Italy UTR5CprMENS1NS2 2014 West Nile virus strain ANS2BNS3NS4A2KN Italy/2014/Pavia5, S4BNS5UTR3 complete genome AJR27894 3434 2 Italy UTR5CprMENS1NS2 2014 West Nile virus strain ANS2BNS3NS4A2KN Italy/2014/Verona/35.1, S4BNS5UTR3 complete genome AJR27895 3434 2 Italy CprMENS1NS2ANS2 2014 West Nile virus strain BNS3NS4A2KNS4BN Italy/2014/Verona/35.2, S5UTR3 complete genome AAQ00999 3433 1A Tunisia UTR5CprMENS1NS2 1997 West Nile virus strain ANS2BNS3NS4A2KN PaH001 polyprotein S4BNS5UTR3 (pol) gene, complete cds AAQ55854 3433 1A USA UTR5CprMENS1NS2 2002 West Nile virus strain ANS2BNS3NS4A2KN TVP 8533 complete S4BNS5 genome AKH14860 3434 2 Austria UTR5CprMENS1NS2 2014 West Nile virus strain ANS2BNS3NS4A2KN blood S4BNS5UTR3 donor/Vienna/2014, complete genome

TABLE12 EEEVNucleic Acid Sequences Antigen Nucleic Acid Sequence SEQ ID NO: SE_EEEv_FL9 ORF: 48 3-939_E2E1 ATGGATTTGGACACTCATTTCACCCAGTATAAGTTGGCACGCCCGT EEE, USA ATATTGCTGATTGCCCTAACTGTGGGCATAGTCGGTGCGACAGCCC isolate TATAGCTATAGAAGAAGTCAGAGGGGATGCGCACGCAGGAGTCATC CGCATCCAGACATCAGCTATGTTCGGTCTGAAGACGGATGGAGTCG ATTTGGCCTACATGAGTTTCATGAACGGCAAAACGCAGAAATCAAT AAAGATCGACAACCTGCATGTGCGCACCTCAGCCCCTTGTTCCCTC GTGTCGCACCACGGCTATTACATCCTGGCTCAATGCCCACCAGGGG ACACGGTTACAGTTGGGTTTCACGACGGGCCTAACCGCCATACGTG CACAGTTGCCCATAAGGTAGAATTCAGGCCAGTGGGTAGAGAGAAA TACCGTCACCCACCTGAACATGGAGTTGAATTACCGTGTAACCGTT ACACTCACAAGCGTGCAGACCAAGGACACTATGTTGAGATGCATCA ACCCGGGCTAGTTGCCGACCACTCTCTCCTTAGCATCCACAGTGCC AAGGTGAAAATTACGGTACCGAGCGGCGCCCAAGTGAAATACTACT GCAAGTGCCCAGATGTACGAGAGGGAATTACCAGCAGCGACCATAC AACCACCTGCACGGATGTCAAACAATGCAGGGCTTACCTGATTGAC AACAAAAAATGGGTGTACAACTCTGGAAGACTGCCTCGAGGAGAGG GCGACACTTTTAAAGGAAAACTTCATGTGCCCTTTGTGCCTGTTAA GGCCAAGTGCATCGCCACGCTGGCACCGGAGCCTCTAGTTGAGCAC AAACACCGCACCCTGATTTTACACCTGCACCCGGACCATCCGACCT TGCTGACGACCAGGTCACTTGGAAGTGATGCAAATCCAACTCGACA ATGGATTGAGCGACCAACAACTGTCAATTTCACAGTCACCGGAGAA GGGTTGGAGTATACCTGGGGAAACCATCCACCAAAAAGAGTATGGG CTCAAGAGTCAGGAGAAGGGAATCCACATGGATGGCCGCACGAAGT GGTAGTCTATTACTACAACAGGTACCCGTTAACCACAATTATCGGA TTATGCACCTGTGTGGCTATCATTATGGTCTCTTGTGTCACATCCG TGTGGCTCCTTTGCAGGACTCGCAATCTTTGCATAACCCCGTATAA ACTAGCCCCGAACGCTCAAGTCCCAATACTCCTGGCGTTACTCTGC TGCATTAAGCCGACGAGGGCATACGAACACACAGCAGTGATGCCGA ACAAGGTGGGGATCCCGTATAAAGCTTTAGTCGAACGCCCAGGTTA TGCACCCGTTCACCTACAGATACAGCTGGTTAATACCAGGATAATT CCATCAACCAACCTGGAGTACATCACCTGCAAGTACAAGACAAAAG TGCCTTCTCCAGTAGTGAAATGCTGCGGTGCCACTCAATGTACCTC CAAACCCCATCCTGACTATCAGTGTCAGGTGTTTACAGGTGTTTAC CCATTCATGTGGGGAGGAGCCTACTGCTTCTGTGACACCGAAAACA CCCAGATGAGCGAGGCGTATGTAGAGCGCTCGGAAGAGTGCTCTAT TGACCACGCAAAAGCTTATAAAGTACACACAGGCACTGTTCAGGCA ATGGTGAACATAACTTATGGGAGCGTCAGCTGGAGATCTGCAGATG TCTACGTCAATGGTGAAACTCCCGCGAAAATAGGAGATGCCAAACT CATCATAGGTCCACTGTCATCTGCGTGGTCCCCATTCGATAACAAG GTGGTGGTTTATGGGCATGAAGTGTATAATTACGACTTTCCTGAGT ACGGCACCGGCAAAGCAGGCTCTTTTGGAGACCTGCAATCACGCAC ATCAACCAGCAACGATCTGTACGCAAACACCAACTTGAAGCTACAA CGACCCCAGGCTGGTATCGTACACACACCTTTCACCCAGGCGCCCT CTGGCTTCGAACGATGGAAAAGGGACAAAGGGGCACCGTTGAACGA CGTAGCCCCGTTTGGCTGTTCGATTGCCCTGGAGCCGCTCCGTGCA GAAAATTGTGCAGTGGGAAGCATCCCTATATCTATAGATATACCCG ATGCGGCTTTCACCAGAATATCTGAAACACCGACAGTCTCAGACCT GGAATGCAAAATTACGGAGTGTACTTATGCCTCCGACTTCGGTGGT ATAGCCACCGTTGCCTACAAATCCAGTAAAGCAGGAAACTGTCCAA TTCATTCTCCATCAGGTGTTGCAGTTATTAAAGAGAATGACGTTAC TCTTGCTGAGAGCGGATCATTTACATTCCACTTCTCCACTGCAAAC ATCCATCCTGCTTTTAAGCTGCAGGTCTGCACTAGTGCAGTTACCT GCAAAGGAGATTGTAAGCCACCGAAAGACCACATCGTCGATTATCC AGCACAACATACCGAATCCTTTACGTCGGCGATATCCGCCACCGCG TGGTCGTGGCTAAAAGTGCTGGTAGGAGGAACATCAGCATTCATTG TTCTGGGGCTTATTGCTACAGCAGTGGTTGCCCTAGTTTTGTTCTT CCATAGACAT SE_EEEv_FL9 ORF: 49 3-939_E2E1_SP ATGCAGAGAGTTGTGTTTGTCGTGCTATTGCTTTTGGTGGCCCCAG EEE, USA CTTACAGCGATTTGGACACTCATTTCACCCAGTATAAGTTGGCACG isolate CCCGTATATTGCTGATTGCCCTAACTGTGGGCATAGTCGGTGCGAC AGCCCTATAGCTATAGAAGAAGTCAGAGGGGATGCGCACGCAGGAG TCATCCGCATCCAGACATCAGCTATGTTCGGTCTGAAGACGGATGG AGTCGATTTGGCCTACATGAGTTTCATGAACGGCAAAACGCAGAAA TCAATAAAGATCGACAACCTGCATGTGCGCACCTCAGCCCCTTGTT CCCTCGTGTCGCACCACGGCTATTACATCCTGGCTCAATGCCCACC AGGGGACACGGTTACAGTTGGGTTTCACGACGGGCCTAACCGCCAT ACGTGCACAGTTGCCCATAAGGTAGAATTCAGGCCAGTGGGTAGAG AGAAATACCGTCACCCACCTGAACATGGAGTTGAATTACCGTGTAA CCGTTACACTCACAAGCGTGCAGACCAAGGACACTATGTTGAGATG CATCAACCCGGGCTAGTTGCCGACCACTCTCTCCTTAGCATCCACA GTGCCAAGGTGAAAATTACGGTACCGAGCGGCGCCCAAGTGAAATA CTACTGCAAGTGCCCAGATGTACGAGAGGGAATTACCAGCAGCGAC CATACAACCACCTGCACGGATGTCAAACAATGCAGGGCTTACCTGA TTGACAACAAAAAATGGGTGTACAACTCTGGAAGACTGCCTCGAGG AGAGGGCGACACTTTTAAAGGAAAACTTCATGTGCCCTTTGTGCCT GTTAAGGCCAAGTGCATCGCCACGCTGGCACCGGAGCCTCTAGTTG AGCACAAACACCGCACCCTGATTTTACACCTGCACCCGGACCATCC GACCTTGCTGACGACCAGGTCACTTGGAAGTGATGCAAATCCAACT CGACAATGGATTGAGCGACCAACAACTGTCAATTTCACAGTCACCG GAGAAGGGTTGGAGTATACCTGGGGAAACCATCCACCAAAAAGAGT ATGGGCTCAAGAGTCAGGAGAAGGGAATCCACATGGATGGCCGCAC GAAGTGGTAGTCTATTACTACAACAGGTACCCGTTAACCACAATTA TCGGATTATGCACCTGTGTGGCTATCATTATGGTCTCTTGTGTCAC ATCCGTGTGGCTCCTTTGCAGGACTCGCAATCTTTGCATAACCCCG TATAAACTAGCCCCGAACGCTCAAGTCCCAATACTCCTGGCGTTAC TCTGCTGCATTAAGCCGACGAGGGCATACGAACACACAGCAGTGAT GCCGAACAAGGTGGGGATCCCGTATAAAGCTTTAGTCGAACGCCCA GGTTATGCACCCGTTCACCTACAGATACAGCTGGTTAATACCAGGA TAATTCCATCAACCAACCTGGAGTACATCACCTGCAAGTACAAGAC AAAAGTGCCTTCTCCAGTAGTGAAATGCTGCGGTGCCACTCAATGT ACCTCCAAACCCCATCCTGACTATCAGTGTCAGGTGTTTACAGGTG TTTACCCATTCATGTGGGGAGGAGCCTACTGCTTCTGTGACACCGA AAACACCCAGATGAGCGAGGCGTATGTAGAGCGCTCGGAAGAGTGC TCTATTGACCACGCAAAAGCTTATAAAGTACACACAGGCACTGTTC AGGCAATGGTGAACATAACTTATGGGAGCGTCAGCTGGAGATCTGC AGATGTCTACGTCAATGGTGAAACTCCCGCGAAAATAGGAGATGCC AAACTCATCATAGGTCCACTGTCATCTGCGTGGTCCCCATTCGATA ACAAGGTGGTGGTTTATGGGCATGAAGTGTATAATTACGACTTTCC TGAGTACGGCACCGGCAAAGCAGGCTCTTTTGGAGACCTGCAATCA CGCACATCAACCAGCAACGATCTGTACGCAAACACCAACTTGAAGC TACAACGACCCCAGGCTGGTATCGTACACACACCTTTCACCCAGGC GCCCTCTGGCTTCGAACGATGGAAAAGGGACAAAGGGGCACCGTTG AACGACGTAGCCCCGTTTGGCTGTTCGATTGCCCTGGAGCCGCTCC GTGCAGAAAATTGTGCAGTGGGAAGCATCCCTATATCTATAGATAT ACCCGATGCGGCTTTCACCAGAATATCTGAAACACCGACAGTCTCA GACCTGGAATGCAAAATTACGGAGTGTACTTATGCCTCCGACTTCG GTGGTATAGCCACCGTTGCCTACAAATCCAGTAAAGCAGGAAACTG TCCAATTCATTCTCCATCAGGTGTTGCAGTTATTAAAGAGAATGAC GTTACTCTTGCTGAGAGCGGATCATTTACATTCCACTTCTCCACTG CAAACATCCATCCTGCTTTTAAGCTGCAGGTCTGCACTAGTGCAGT TACCTGCAAAGGAGATTGTAAGCCACCGAAAGACCACATCGTCGAT TATCCAGCACAACATACCGAATCCTTTACGTCGGCGATATCCGCCA CCGCGTGGTCGTGGCTAAAAGTGCTGGTAGGAGGAACATCAGCATT CATTGTTCTGGGGCTTATTGCTACAGCAGTGGTTGCCCTAGTTTTG TTCTTCCATAGACAT EEEmRNASequences SE_EEEv_FL9 ORF: 50 SE_EEEv_FL9 AUGGAUUUGGACACUCAUUUCACCCAGUAUAAGUUGGCACGCCCGU 3-939_E2E1 AUAUUGCUGAUUGCCCUAACUGUGGGCAUAGUCGGUGCGACAGCCC EEE, USA UAUAGCUAUAGAAGAAGUCAGAGGGGAUGCGCACGCAGGAGUCAUC isolate CGCAUCCAGACAUCAGCUAUGUUCGGUCUGAAGACGGAUGGAGUCG AUUUGGCCUACAUGAGUUUCAUGAACGGCAAAACGCAGAAAUCAAU AAAGAUCGACAACCUGCAUGUGCGCACCUCAGCCCCUUGUUCCCUC GUGUCGCACCACGGCUAUUACAUCCUGGCUCAAUGCCCACCAGGGG ACACGGUUACAGUUGGGUUUCACGACGGGCCUAACCGCCAUACGUG CACAGUUGCCCAUAAGGUAGAAUUCAGGCCAGUGGGUAGAGAGAAA UACCGUCACCCACCUGAACAUGGAGUUGAAUUACCGUGUAACCGUU ACACUCACAAGCGUGCAGACCAAGGACACUAUGUUGAGAUGCAUCA ACCCGGGCUAGUUGCCGACCACUCUCUCCUUAGCAUCCACAGUGCC AAGGUGAAAAUUACGGUACCGAGCGGCGCCCAAGUGAAAUACUACU GCAAGUGCCCAGAUGUACGAGAGGGAAUUACCAGCAGCGACCAUAC AACCACCUGCACGGAUGUCAAACAAUGCAGGGCUUACCUGAUUGAC AACAAAAAAUGGGUGUACAACUCUGGAAGACUGCCUCGAGGAGAGG GCGACACUUUUAAAGGAAAACUUCAUGUGCCCUUUGUGCCUGUUAA GGCCAAGUGCAUCGCCACGCUGGCACCGGAGCCUCUAGUUGAGCAC AAACACCGCACCCUGAUUUUACACCUGCACCCGGACCAUCCGACCU UGCUGACGACCAGGUCACUUGGAAGUGAUGCAAAUCCAACUCGACA AUGGAUUGAGCGACCAACAACUGUCAAUUUCACAGUCACCGGAGAA GGGUUGGAGUAUACCUGGGGAAACCAUCCACCAAAAAGAGUAUGGG CUCAAGAGUCAGGAGAAGGGAAUCCACAUGGAUGGCCGCACGAAGU GGUAGUCUAUUACUACAACAGGUACCCGUUAACCACAAUUAUCGGA UUAUGCACCUGUGUGGCUAUCAUUAUGGUCUCUUGUGUCACAUCCG UGUGGCUCCUUUGCAGGACUCGCAAUCUUUGCAUAACCCCGUAUAA ACUAGCCCCGAACGCUCAAGUCCCAAUACUCCUGGCGUUACUCUGC UGCAUUAAGCCGACGAGGGCAUACGAACACACAGCAGUGAUGCCGA ACAAGGUGGGGAUCCCGUAUAAAGCUUUAGUCGAACGCCCAGGUUA UGCACCCGUUCACCUACAGAUACAGCUGGUUAAUACCAGGAUAAUU CCAUCAACCAACCUGGAGUACAUCACCUGCAAGUACAAGACAAAAG UGCCUUCUCCAGUAGUGAAAUGCUGCGGUGCCACUCAAUGUACCUC CAAACCCCAUCCUGACUAUCAGUGUCAGGUGUUUACAGGUGUUUAC CCAUUCAUGUGGGGAGGAGCCUACUGCUUCUGUGACACCGAAAACA CCCAGAUGAGCGAGGCGUAUGUAGAGCGCUCGGAAGAGUGCUCUAU UGACCACGCAAAAGCUUAUAAAGUACACACAGGCACUGUUCAGGCA AUGGUGAACAUAACUUAUGGGAGCGUCAGCUGGAGAUCUGCAGAUG UCUACGUCAAUGGUGAAACUCCCGCGAAAAUAGGAGAUGCCAAACU CAUCAUAGGUCCACUGUCAUCUGCGUGGUCCCCAUUCGAUAACAAG GUGGUGGUUUAUGGGCAUGAAGUGUAUAAUUACGACUUUCCUGAGU ACGGCACCGGCAAAGCAGGCUCUUUUGGAGACCUGCAAUCACGCAC AUCAACCAGCAACGAUCUGUACGCAAACACCAACUUGAAGCUACAA CGACCCCAGGCUGGUAUCGUACACACACCUUUCACCCAGGCGCCCU CUGGCUUCGAACGAUGGAAAAGGGACAAAGGGGCACCGUUGAACGA CGUAGCCCCGUUUGGCUGUUCGAUUGCCCUGGAGCCGCUCCGUGCA GAAAAUUGUGCAGUGGGAAGCAUCCCUAUAUCUAUAGAUAUACCCG AUGCGGCUUUCACCAGAAUAUCUGAAACACCGACAGUCUCAGACCU GGAAUGCAAAAUUACGGAGUGUACUUAUGCCUCCGACUUCGGUGGU AUAGCCACCGUUGCCUACAAAUCCAGUAAAGCAGGAAACUGUCCAA UUCAUUCUCCAUCAGGUGUUGCAGUUAUUAAAGAGAAUGACGUUAC UCUUGCUGAGAGCGGAUCAUUUACAUUCCACUUCUCCACUGCAAAC AUCCAUCCUGCUUUUAAGCUGCAGGUCUGCACUAGUGCAGUUACCU GCAAAGGAGAUUGUAAGCCACCGAAAGACCACAUCGUCGAUUAUCC AGCACAACAUACCGAAUCCUUUACGUCGGCGAUAUCCGCCACCGCG UGGUCGUGGCUAAAAGUGCUGGUAGGAGGAACAUCAGCAUUCAUUG UUCUGGGGCUUAUUGCUACAGCAGUGGUUGCCCUAGUUUUGUUCUU CCAUAGACAU SE_EEEv_FL9 ORF: 51 SE_EEEv_FL9 AUGCAGAGAGUUGUGUUUGUCGUGCUAUUGCUUUUGGUGGCCCCAG 3-939_E2E1_SP CUUACAGCGAUUUGGACACUCAUUUCACCCAGUAUAAGUUGGCACG EEE, USA CCCGUAUAUUGCUGAUUGCCCUAACUGUGGGCAUAGUCGGUGCGAC isolate AGCCCUAUAGCUAUAGAAGAAGUCAGAGGGGAUGCGCACGCAGGAG UCAUCCGCAUCCAGACAUCAGCUAUGUUCGGUCUGAAGACGGAUGG AGUCGAUUUGGCCUACAUGAGUUUCAUGAACGGCAAAACGCAGAAA UCAAUAAAGAUCGACAACCUGCAUGUGCGCACCUCAGCCCCUUGUU CCCUCGUGUCGCACCACGGCUAUUACAUCCUGGCUCAAUGCCCACC AGGGGACACGGUUACAGUUGGGUUUCACGACGGGCCUAACCGCCAU ACGUGCACAGUUGCCCAUAAGGUAGAAUUCAGGCCAGUGGGUAGAG AGAAAUACCGUCACCCACCUGAACAUGGAGUUGAAUUACCGUGUAA CCGUUACACUCACAAGCGUGCAGACCAAGGACACUAUGUUGAGAUG CAUCAACCCGGGCUAGUUGCCGACCACUCUCUCCUUAGCAUCCACA GUGCCAAGGUGAAAAUUACGGUACCGAGCGGCGCCCAAGUGAAAUA CUACUGCAAGUGCCCAGAUGUACGAGAGGGAAUUACCAGCAGCGAC CAUACAACCACCUGCACGGAUGUCAAACAAUGCAGGGCUUACCUGA UUGACAACAAAAAAUGGGUGUACAACUCUGGAAGACUGCCUCGAGG AGAGGGCGACACUUUUAAAGGAAAACUUCAUGUGCCCUUUGUGCCU GUUAAGGCCAAGUGCAUCGCCACGCUGGCACCGGAGCCUCUAGUUG AGCACAAACACCGCACCCUGAUUUUACACCUGCACCCGGACCAUCC GACCUUGCUGACGACCAGGUCACUUGGAAGUGAUGCAAAUCCAACU CGACAAUGGAUUGAGCGACCAACAACUGUCAAUUUCACAGUCACCG GAGAAGGGUUGGAGUAUACCUGGGGAAACCAUCCACCAAAAAGAGU AUGGGCUCAAGAGUCAGGAGAAGGGAAUCCACAUGGAUGGCCGCAC GAAGUGGUAGUCUAUUACUACAACAGGUACCCGUUAACCACAAUUA UCGGAUUAUGCACCUGUGUGGCUAUCAUUAUGGUCUCUUGUGUCAC AUCCGUGUGGCUCCUUUGCAGGACUCGCAAUCUUUGCAUAACCCCG UAUAAACUAGCCCCGAACGCUCAAGUCCCAAUACUCCUGGCGUUAC UCUGCUGCAUUAAGCCGACGAGGGCAUACGAACACACAGCAGUGAU GCCGAACAAGGUGGGGAUCCCGUAUAAAGCUUUAGUCGAACGCCCA GGUUAUGCACCCGUUCACCUACAGAUACAGCUGGUUAAUACCAGGA UAAUUCCAUCAACCAACCUGGAGUACAUCACCUGCAAGUACAAGAC AAAAGUGCCUUCUCCAGUAGUGAAAUGCUGCGGUGCCACUCAAUGU ACCUCCAAACCCCAUCCUGACUAUCAGUGUCAGGUGUUUACAGGUG UUUACCCAUUCAUGUGGGGAGGAGCCUACUGCUUCUGUGACACCGA AAACACCCAGAUGAGCGAGGCGUAUGUAGAGCGCUCGGAAGAGUGC UCUAUUGACCACGCAAAAGCUUAUAAAGUACACACAGGCACUGUUC AGGCAAUGGUGAACAUAACUUAUGGGAGCGUCAGCUGGAGAUCUGC AGAUGUCUACGUCAAUGGUGAAACUCCCGCGAAAAUAGGAGAUGCC AAACUCAUCAUAGGUCCACUGUCAUCUGCGUGGUCCCCAUUCGAUA ACAAGGUGGUGGUUUAUGGGCAUGAAGUGUAUAAUUACGACUUUCC UGAGUACGGCACCGGCAAAGCAGGCUCUUUUGGAGACCUGCAAUCA CGCACAUCAACCAGCAACGAUCUGUACGCAAACACCAACUUGAAGC UACAACGACCCCAGGCUGGUAUCGUACACACACCUUUCACCCAGGC GCCCUCUGGCUUCGAACGAUGGAAAAGGGACAAAGGGGCACCGUUG AACGACGUAGCCCCGUUUGGCUGUUCGAUUGCCCUGGAGCCGCUCC GUGCAGAAAAUUGUGCAGUGGGAAGCAUCCCUAUAUCUAUAGAUAU ACCCGAUGCGGCUUUCACCAGAAUAUCUGAAACACCGACAGUCUCA GACCUGGAAUGCAAAAUUACGGAGUGUACUUAUGCCUCCGACUUCG GUGGUAUAGCCACCGUUGCCUACAAAUCCAGUAAAGCAGGAAACUG UCCAAUUCAUUCUCCAUCAGGUGUUGCAGUUAUUAAAGAGAAUGAC GUUACUCUUGCUGAGAGCGGAUCAUUUACAUUCCACUUCUCCACUG CAAACAUCCAUCCUGCUUUUAAGCUGCAGGUCUGCACUAGUGCAGU UACCUGCAAAGGAGAUUGUAAGCCACCGAAAGACCACAUCGUCGAU UAUCCAGCACAACAUACCGAAUCCUUUACGUCGGCGAUAUCCGCCA CCGCGUGGUCGUGGCUAAAAGUGCUGGUAGGAGGAACAUCAGCAUU CAUUGUUCUGGGGCUUAUUGCUACAGCAGUGGUUGCCCUAGUUUUG UUCUUCCAUAGACAU

For all sequences in Table 12:

5′UTR: DNA (SEQ ID NO: 40) TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAA ATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACC 3′UTR: DNA (SEQ ID NO: 41) TGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTC CCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCGTGGTCTTTGAA TAAAGTCTGAGTGGGCGGC 5′UTR: mRNA (SEQ ID NO: 42) UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAA AUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACC 3′UTR: mRNA (SEQ ID NO: 43) UGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUC CCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCGUGGUCUUUGAA UAAAGUCUGAGUGGGCGGC

TABLE 13 EEEV Amino Acid Sequences Antigen Amino Acid Sequence SEQ ID NO: SE_EEEv_FL93- MDLDTHFTQYKLARPYIADCPNCGHSRCDSPIAIEEVRGDAHAGVI 52 939_E2E1 RIQTSAMFGEKTDGVDLAYMSFMNGKTQKSIKIDNEHVRTSAPCSL VSHHGYYILAQCPPGDTVTVGFHDGPNRHTCTVAHKVEFRPVGREK YRHPPEHGVELPCNRYTHKRADQGHYVEMHQPGLVADHSLLSIHSA KVKITVPSGAQVKYYCKCPDVREGITSSDHTTTCTDVKQCRAYLID NKKWVYNSGREPRGEGDTPKGKEHVPFVPVKAKCIATLAPEPLVEH KHRTLIEHLHPDHPTLETTRSLGSDANPTRQWIERPTTVNFTVTGE GLEYTWGNHPPKRVWAQESGEGNPHGWPHEVVVYYYNRYPLTTIIG LCTCVAIIMVSCVTSINVWELCRTRNLCITPYKLAPNAQVPILLAL LCCIKPTRAYEHTAVMPNKVGIPYKALVERPGYAPVHLQIQLVNTR IIPSTNLEYITCKYKTKVPSPVVKCCGATQCTSKPHPDYQCQVFTG VYPFMWGGAYCFCDTENTQMSEAYVERSEECSIDHAKAYKVHTGTV QAMVNITYGSINVSWRSADVYVNGETPAKIGDAKLIIGPLSSAWSP FDNKVVVYGHEVYNYDFPEYGTGKAGSFGDLQSRTSTSNDLYANTN EKLQRPQAGIVHTPFTQAPSGFERWKRDKGAPENDVAPFGCSIALE PLRAENCAVGSIPISIDIPDAAFTRISETPTVSDLECKITECTYAS DFGGIATVAYKSSKAGNCPIHSPSGVAVIKENDVTLAESGSFTFHF STANIHPAFKLQVCTSAVTCKGDCKPPKDHIVDYPAQHTESFTSAI SATAWSWEKVLVGGTSAFIVEGLIATAVVALVEFFHRH SE_EEEv_FL93- MQRVVFVVELLEVAPAYSDLDTHFTQYKLARPYIADCPNCGHSRCD 53 939-E2E1-SP SPIAIEEVRGDAHAGVIRIQTSAMFGLKTDGVDLAYMSFMNGKTQK SIKIDNEHVRTSAPCSLVSHHGYYILAQCPPGDTVTVGFHDGPNRH TCTVAHKVEFRPVGREKYRHPPEHGVELPCNRYTHKRADQGHYVEM HQPGLVADHSLLSIHSAKVKITVPSGAQVKYYCKCPDVREGITSSD HTTTCTDVKQCRAYLIDNKKWVYNSGREPRGEGDTPKGKEHVPFVP VKAKCIATLAPEPLVEHKHRTLIEHLHPDHPTLETTRSLGSDANPT RQWIERPTTVNFTVTGEGLEYTWGNHPPKRVWAQESGEGNPHGWPH EVVVYYYNRYPETTIIGLCTCVAIIMVSCVTSINVWELCRTRNLCI TPYKLAPNAQVPILLALLCCIKPTRAYEHTAVMPNKVGIPYKALVE RPGYAPVHLQIQLVNTRIIPSTNLEYITCKYKTKVPSPVVKCCGAT QCTSKPHPDYQCQVFTGVYPFMWGGAYCFCDTENTQMSEAYVERSE ECSIDHAKAYKVHTGTVQAMVNITYGSINVSWRSADVYVNGETPAK IGDAKLIIGPLSSAWSPFDNKVVVYGHEVYNYDPPEYGTGKAGSFG DEQSRTSTSNDLYANTNEKLQRPQAGIVHTPFTQAPSGFERWKRDK GAPLNDVAPFGCSIALEPLRAENCAVGSIPISIDIPDAAFTRISET PTVSDLECKITECTYASDPGGIATVAYKSSKAGNCPIHSPSGVAVI KENDVTLAESGSFTFHFSTANIHPAFKLQVCTSAVTCKGDCKPPKD HIVDYPAQHTESFTSAISATAWSWEKVLVGGTSAFIVEGLIATAVV ALVLFFHRH SE_EEEv_FL93- MDLDTHFTQYKLARPYIADCPNCGHSRCDSPIAIEEVRGDAHAGVI 54 939_E2E1 RIQTSAMFGLKTDGVDLAYMSFMNGKTQKSIKIDNLHVRTSAPCSL VSHHGYYILAQCPPGDTVTVGFHDGPNRHTCTVAHKVEFRPVGREK YRHPPEHGVELPCNRYTHKRADQGHYVEMHQPGLVADHSLLSIHSA KVKITVPSGAQVKYYCKCPDVREGITSSDHTTTCTDVKQCRAYLID NKKWVYNSGRLPRGEGDTFKGKLHVPFVPVKAKCIATLAPEPLVEH KHRTLILHLHPDHPTLLTTRSLGSDANPTRQWIERPTTVNFTVTGE GLEYTWGNHPPKRVWAQESGEGNPHGWPHEVVVYYYNRYPLTTIIG LCTCVAIIMVSCVTSINVWLLCRTRNLCITPYKLAPNAQVPILLAL LCCIKPTRAYEHTAVMPNKVGIPYKALVERPGYAPVHLQIQLVNTR IIPSTNLEYITCKYKTKVPSPVVKCCGATQCTSKPHPDYQCQVFTG VYPFMWGGAYCFCDTENTQMSEAYVERSEECSIDHAKAYKVHTGTV QAMVNITYGSINVSWRSADVYVNGETPAKIGDAKLIIGPLSSAWSP FDNKVVVYGHEVYNYDFPEYGTGKAGSFGDLQSRTSTSNDLYANTN LKLQRPQAGIVHTPFTQAPSGFERWKRDKGAPLNDVAPFGCSIALE PLRAENCAVGSIPISIDIPDAAFTRISETPTVSDLECKITECTYAS DFGGIATVAYKSSKAGNCPIHSPSGVAVIKENDVTLAESGSFTFHF STANIHPAFKLQVCTSAVTCKGDCKPPKDHIVDYPAQHTESFTSAI SATAWSWLKVLVGGTSAFIVLGLIATAVVALVLFFHRH

TABLE 14 EEEV strains/isolates, EEEV Amino Acid Sequences GenBank Type Virus Name Accession non- non-structural polyprotein precursor P1234 [Eastern equine AHL83632.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83782.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83714.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83684.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83756.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83630.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83730.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83814.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83784.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83758.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83732.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83656.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83762.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83820.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83770.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83690.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83822.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83806.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83744.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83818.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83776.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83750.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83704.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83666.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83778.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83648.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83738.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83736.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83662.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83640.1 structural encephalitis virus] non- nonstructural polyprotein [Eastern equine encephalitis virus] ADW86066.1 structural non- non-structural polyprotein precursor P1234 [Eastern equine AHL83788.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83786.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83752.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83664.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83660.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83774.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83728.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83702.1 structural encephalitis virus] non- nonstructural protein [Eastern equine encephalitis virus] ADW86060.1 structural non- nonstructural protein [Eastern equine encephalitis virus] ADW86062.1 structural non- nonstructural protein [Eastern equine encephalitis virus] AAT96377.1 structural non- non-structural polyprotein precursor P1234 [Eastern equine AHL83734.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83678.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83798.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83674.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83634.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83688.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83654.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83724.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83686.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83638.1 structural encephalitis virus] non- nonstructural protein [Eastern equine encephalitis virus] ADW86064.1 structural non- non-structural polyprotein precursor P1234 [Eastern equine AHL83652.1 structural encephalitis virus] non- nonstructural polyprotein [Eastern equine encephalitis virus] ADW86058.1 structural non- non-structural polyprotein precursor P1234 [Eastern equine AHL83740.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83694.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83718.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83710.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83708.1 structural encephalitis virus] non- nonstructural protein [Eastern equine encephalitis virus] AAT96379.1 structural non- non-structural polyprotein precursor P1234 [Eastern equine AHL83792.1 structural encephalitis virus] non- nonstructural polyprotein [Eastern equine encephalitis virus] ADW86068.1 structural non- non-structural polyprotein precursor P1234 [Eastern equine AHL83760.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83790.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHW48355.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83726.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83642.1 structural encephalitis virus] non- nonstructural protein [Eastern equine encephalitis virus] AGI74964.1 structural non- non-structural polyprotein precursor P1234 [Eastern equine AHL83768.1 structural encephalitis virus] non- Non-structural polyprotein; Polyprotein nsP1234; Q4QXJ8.2 structural ContainsP123; Non-structural protein 1; Non-structural protein 2; Non-structural protein 3; nsP4 non- non-structural polyprotein precursor P1234 [Eastern equine AHL83742.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83646.1 structural encephalitis virus] non- non-structural polyprotein precursor P1234 [Eastern equine AHL83780.1 structural encephalitis virus] non- non-structural polyprotein precursor [Eastern equine ABQ63085.1 structural encephalitis virus] non- nonstructural protein [Eastern equine encephalitis virus] AAU95734.1 structural non- nonstructural polyprotein [Eastern equine encephalitis virus] ABL84686.1 structural non- non-structural polyprotein precursor P1234 [Eastern equine AHL83706.1 structural encephalitis virus] non- nonstructural polyprotein [Eastern equine encephalitis virus] AAC53734.1 structural non- nonstructural polyprotein p1234 [Eastern equine encephalitis NP_632021.2 structural virus] structural structural polyprotein [Eastern equine encephalitis virus] AHL83633.1 structural Structural polyprotein; p130; Contains: Capsid protein; Coat Q4QXJ7.1 protein p62; E3 protein; E2 envelope glycoprotein; 6K protein; E1 envelope glycoprotein; structural structural polyprotein [Eastern equine encephalitis virus] AHL83719.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83785.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83759.1 structural structural polyprotein [Eastern equine encephalitis virus] AAT96378.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83797.1 structural structural polyprotein [Eastern equine encephalitis virus] ADB08660.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83815.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83761.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83685.1 structural structural polyprotein [Eastern equine encephalitis virus] ADW86011.2 structural structural polyprotein [Eastern equine encephalitis virus] ADW86009.2 structural structural polyprotein [Eastern equine encephalitis virus] ADW86069.1 structural structural polyprotein [Eastern equine encephalitis virus] ADW86067.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83819.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83769.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83751.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83737.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83679.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83639.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83809.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83793.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83707.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83655.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83649.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83781.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83757.1 structural structural polyprotein [Eastern equine encephalitis virus] ADW86015.2 structural structural polyprotein [Eastern equine encephalitis virus] ADW86005.2 structural structural polyprotein [Eastern equine encephalitis virus] AHL83741.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83727.1 structural structural polyprotein [Eastern equine encephalitis virus] ADW86019.2 structural structural polyprotein [Eastern equine encephalitis virus] AAC53755.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83753.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83735.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83729.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83687.1 structural structural polyprotein [Eastern equine encephalitis virus] ADW86018.2 structural structural polyprotein [Eastern equine encephalitis virus] ADW86059.1 structural structural polyprotein precursor p130 [Eastern equine ABQ63086.1 encephalitis virus] structural structural polyprotein [Eastern equine encephalitis virus] AAF04799.1 structural structural polyprotein [Eastern equine encephalitis virus] AAC53735.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83743.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83667.1 structural structural polyprotein [Eastern equine encephalitis virus] AAC53762.1 structural structural polyprotein [Eastern equine encephalitis virus] AAC53759.1 structural structural polyprotein [Eastern equine encephalitis virus] AAF04793.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83661.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83653.1 structural structural polyprotein [Eastern equine encephalitis virus] AAC53761.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83695.1 structural structural polyprotein [Eastern equine encephalitis virus] AAC53760.1 structural structural polyprotein [Eastern equine encephalitis virus] AAF04794.1 structural structural polyprotein [Eastern equine encephalitis virus] AHL83791.1 structural structural polyprotein [Eastern equine encephalitis virus] AAC53756.1 structural structural polyprotein [Eastern equine encephalitis virus] AAC53758.1 structural polyprotein [Eastern equine encephalitis virus] AAA67908.1 structural p130 [Eastern equine encephalitis virus] ALE15082.1 structural structural proteins polyprotein [Eastern equine encephalitis AAA02897.1 virus] structural structural polyprotein [Eastern equine encephalitis virus] AAC53757.1 structural structural polyprotein [Eastern equine encephalitis virus] ACY66806.1 structural structural polyprotein [Eastern equine encephalitis virus] AAF04795.1 structural structural polyprotein [Eastern equine encephalitis virus] AAF04800.1 structural structural polyprotein [Eastern equine encephalitis virus] AAF04792.1 structural structural polyprotein [Eastern equine encephalitis virus] AAF04798.1 structural precursor of structural proteins [Eastern equine encephalitis NP_632022.1 virus] structural structural polyprotein [Eastern equine encephalitis virus] AAF04797.1 structural structural polyprotein [Eastern equine encephalitis virus] AAF04796.1 structural structural protein [Eastern equine encephalitis virus] AGI74965.1 structural Structural polyprotein; p130; Contains: Capsid protein; Coat P27284.1 protein; p62; E3 protein; E2 envelope glycoprotein; 6K protein; E1 envelope glycoprotein; structural structural polyprotein [Eastern equine encephalitis virus] ADB08676.1 structural Structural polyprotein; p130; Contains Capsid protein; Coat P08768.1 protein; p62; E3 protein; E2 envelope glycoprotein; 6K protein; E1 envelope glycoprotein; structural structural polyprotein [Eastern equine encephalitis virus] ADB08666.1

TABLE 15 SINV Nucleic Acid Sequences Antigen Nucleic Acid Sequence SEQ ID NO: SE_SINV_AR3 ORF: 55 SE_SINV_AR3 ATGGATGTGGACGCTCACTTTACCCAGTATAAATTAGCCCGCCCG 39-BeAr436087_ TATATAGCCGACTGCCCAAATTGCGGTCATGGCAGATGTGACAGT E2E1_SP CCGATCGCTATCGAGGACGTGCGCGGAGACGCGCACGCCGGTTAC SINV, Brazilian ATCCGCATACAGACATCCGCAATGTTCGGCATGAAGTCGGAAGGG isolate GTTGATCTGGCTTACATGAGCTATATGAACGGAAAGGTTTTAAAA GCCATCAAGATTGATAGCCTGTACGTCCGTACATCAGCACCTTGT TCTCTGGTTTCTTATCACGGATACTACCTCCTCGCGCAGTGCCCA CCAGGAGATACTGTAACAGTAGGGTTCCTAGAAGGTACCCACAAG TATATGTGTACAGTAGCGCACCAGGTAAAATTTAACCCGGTGGGG AGAGAGAAATACAGACATCCACCAGAACACGGTGTTGAACTACCC TGCAACAAATACACCCATAAGCGTGCCGATCAAGGCCATTATGTA GAAATGCACCAACCGGGTATGGTCGCCGACCACACTCTGTTGAGC ATGAGCGGCACCAAGGTGAAAGTCACCGCACCGAGCAGTTCGCAA GTGAAATATTACTGCAAATGTCCAGATCTTCAAGAAGGAACTACC AGCGGTGAACACACAACAACATGTACCGATGTTAAGCAATGCCGA GCGTATCTGATTGACAACAGGAAGTGGGTGTACAACTCAGGAAAA TTACCTAGAGGAGAAGGCGAAACCTTTAAAGGCAAACTCCATGTA CCATTTGTACCTGTTGCGGCCACCTGCACAGCGACCCTTGCTCCA GAGCCTCTCGTCGAGCACAAGCACCGCTCCCTGATCCTCCACTTG CATCCAGAGCACCCCACGCTATTGACAACAAGGGCGCTCGGAAGT CAAGCACAACCGACTAGGCAATGGATAGACCGCCCAACCACCGTC AACTTCACAGTTACCGGAGAAGGTTTTGAATATACCTGGGGCAAC CATCCCCCGAAAAGAGTATGGGCCCAAGAGTCAGGAGAAGGCAAC CCGCACGGCTGGCCTCACGAGGTGGTAATCTACTACTACAATAGG TACCCGATGACAACTATCGTGGGATTATGCACGTGTGCCGCTATT ATTATGGTGTCATGCGTTACATCTGTATGGCTTTTATGCCGCACC CGTAACCTTTGCATAACACCCTACAGATTGGCACCAAATGCCCAC GTACCTCTCTTAGTAGCGTTGCTGTGCTGCGTTAAACCAACTAGA GCATACGAGCACACGGCTGTGATGTCGAACAAGGTGGGGATCCCT TACAAAGCCCTAGTTGAAAGGCCAGGGTACGCACCCGTACATCTC CAGATCCAGCTGGTAAGCACAAAAATAATCCCTACAGCGAACTTG GAATACATTACCTGTAAATATAAGACTAAGGTGCCTTCCCCAGTA GTAAAATGTTGCGGATCCACCCAATGTACGTCTAAACAATACCCA GACTATCAGTGCCAAGTCTTCACGGGAGTCTACCCATTTATGTGG GGAGGAGCCTACTGTTTCTGTGATACTGAAAATACACAAATGAGC GAAGCGTATGTCGAACGCTCAGAAGAATGCTCAGTGGACCAGGCC AAAGCCTACAAAGTACACACAGGAACGGTGCAGGCTGTAGTTAAT ATCACCTACGGGGGCGTCAGCTGGAGATCTGCCGACGTCTATGTC AACGGAGAGACCCCTGCCAAAATAGGTGATGCTAAACTAACTATA GGCCCTCTCTCTTCCGCATGGTCACCTTTTGACTCTAAAGTCATA GTGTACGGGCACGAGGTGTATAACTATGACTTTCCCGAATACGGT ACCGGCAAAGCCGGTTCGTTCGGAGACTTGCAATCCAGGACACTT ACAAGTAAGGACTTGTACTCTAACACCAATTTGAAACTGCAACGT CCCCAGCCAGGAGTGGTCCACACCCCATACACCCAGGCACCCTCG GGGTTTGAACGTTGGAAGAAAGATCGAGGGGCACCGCTAAACGAC GTCGCTCCTTTTGGATGCAACATAGCCCTGGAACCACTGCGTGCT GAGAACTGTGCGGTGGGGAGTATCCCTCTCTCCATCGACATACCC GATGCCGCTTTTACCAGGATATCGGAGACACCGACTGTCTCCGAT CTGGAATGTAAAATTACTGAGTGCACGTATGCGTCAGATTTTGGG GGAATAGCTACTATGTCGTATAAAGCTAGCAAGGCAGGAAACTGC CCTATTCATTCCCCTTCGGGCATTGCAGTAATTAAAGAGAACGAT GTTATTCTTTCTGGAAGCGGCTCGTTCACGTTCCATTTTTCAACA GCGAGCATCCACCCAGCGTTCAAGATGCAGGTATGCACCAGCGTA GTCACCTGCAAAGGTGACTGCAAGCCACCTAAGGACCATATCGTC GATTACCCAGCCCAGCATACTGAGACGTTTACATCAGCAGTTTCC GCAACTGCATGGTCATGGCTGAAAGTGCTAGTTGGGGGCACATCC GTCTTTATCATTCTTGGGCTAATCGCTACAGCAGTGGTTGCCCTG GTGCTATTCACCCACAAACAC SE_SINV_AR3 ORF: 56 SE_SINV_AR3 ATGCAGAGAGTTGTGTTTGTCGTGCTATTGCTTTTGGTGGCCCCA 39-BeAr436087_ GCTTACAGCGATGTGGACGCTCACTTTACCCAGTATAAATTAGCC E2E1 CGCCCGTATATAGCCGACTGCCCAAATTGCGGTCATGGCAGATGT SINV, Brazilian GACAGTCCGATCGCTATCGAGGACGTGCGCGGAGACGCGCACGCC isolate GGTTACATCCGCATACAGACATCCGCAATGTTCGGCATGAAGTCG GAAGGGGTTGATCTGGCTTACATGAGCTATATGAACGGAAAGGTT TTAAAAGCCATCAAGATTGATAGCCTGTACGTCCGTACATCAGCA CCTTGTTCTCTGGTTTCTTATCACGGATACTACCTCCTCGCGCAG TGCCCACCAGGAGATACTGTAACAGTAGGGTTCCTAGAAGGTACC CACAAGTATATGTGTACAGTAGCGCACCAGGTAAAATTTAACCCG GTGGGGAGAGAGAAATACAGACATCCACCAGAACACGGTGTTGAA CTACCCTGCAACAAATACACCCATAAGCGTGCCGATCAAGGCCAT TATGTAGAAATGCACCAACCGGGTATGGTCGCCGACCACACTCTG TTGAGCATGAGCGGCACCAAGGTGAAAGTCACCGCACCGAGCAGT TCGCAAGTGAAATATTACTGCAAATGTCCAGATCTTCAAGAAGGA ACTACCAGCGGTGAACACACAACAACATGTACCGATGTTAAGCAA TGCCGAGCGTATCTGATTGACAACAGGAAGTGGGTGTACAACTCA GGAAAATTACCTAGAGGAGAAGGCGAAACCTTTAAAGGCAAACTC CATGTACCATTTGTACCTGTTGCGGCCACCTGCACAGCGACCCTT GCTCCAGAGCCTCTCGTCGAGCACAAGCACCGCTCCCTGATCCTC CACTTGCATCCAGAGCACCCCACGCTATTGACAACAAGGGCGCTC GGAAGTCAAGCACAACCGACTAGGCAATGGATAGACCGCCCAACC ACCGTCAACTTCACAGTTACCGGAGAAGGTTTTGAATATACCTGG GGCAACCATCCCCCGAAAAGAGTATGGGCCCAAGAGTCAGGAGAA GGCAACCCGCACGGCTGGCCTCACGAGGTGGTAATCTACTACTAC AATAGGTACCCGATGACAACTATCGTGGGATTATGCACGTGTGCC GCTATTATTATGGTGTCATGCGTTACATCTGTATGGCTTTTATGC CGCACCCGTAACCTTTGCATAACACCCTACAGATTGGCACCAAAT GCCCACGTACCTCTCTTAGTAGCGTTGCTGTGCTGCGTTAAACCA ACTAGAGCATACGAGCACACGGCTGTGATGTCGAACAAGGTGGGG ATCCCTTACAAAGCCCTAGTTGAAAGGCCAGGGTACGCACCCGTA CATCTCCAGATCCAGCTGGTAAGCACAAAAATAATCCCTACAGCG AACTTGGAATACATTACCTGTAAATATAAGACTAAGGTGCCTTCC CCAGTAGTAAAATGTTGCGGATCCACCCAATGTACGTCTAAACAA TACCCAGACTATCAGTGCCAAGTCTTCACGGGAGTCTACCCATTT ATGTGGGGAGGAGCCTACTGTTTCTGTGATACTGAAAATACACAA ATGAGCGAAGCGTATGTCGAACGCTCAGAAGAATGCTCAGTGGAC CAGGCCAAAGCCTACAAAGTACACACAGGAACGGTGCAGGCTGTA GTTAATATCACCTACGGGGGCGTCAGCTGGAGATCTGCCGACGTC TATGTCAACGGAGAGACCCCTGCCAAAATAGGTGATGCTAAACTA ACTATAGGCCCTCTCTCTTCCGCATGGTCACCTTTTGACTCTAAA GTCATAGTGTACGGGCACGAGGTGTATAACTATGACTTTCCCGAA TACGGTACCGGCAAAGCCGGTTCGTTCGGAGACTTGCAATCCAGG ACACTTACAAGTAAGGACTTGTACTCTAACACCAATTTGAAACTG CAACGTCCCCAGCCAGGAGTGGTCCACACCCCATACACCCAGGCA CCCTCGGGGTTTGAACGTTGGAAGAAAGATCGAGGGGCACCGCTA AACGACGTCGCTCCTTTTGGATGCAACATAGCCCTGGAACCACTG CGTGCTGAGAACTGTGCGGTGGGGAGTATCCCTCTCTCCATCGAC ATACCCGATGCCGCTTTTACCAGGATATCGGAGACACCGACTGTC TCCGATCTGGAATGTAAAATTACTGAGTGCACGTATGCGTCAGAT TTTGGGGGAATAGCTACTATGTCGTATAAAGCTAGCAAGGCAGGA AACTGCCCTATTCATTCCCCTTCGGGCATTGCAGTAATTAAAGAG AACGATGTTATTCTTTCTGGAAGCGGCTCGTTCACGTTCCATTTT TCAACAGCGAGCATCCACCCAGCGTTCAAGATGCAGGTATGCACC AGCGTAGTCACCTGCAAAGGTGACTGCAAGCCACCTAAGGACCAT ATCGTCGATTACCCAGCCCAGCATACTGAGACGTTTACATCAGCA GTTTCCGCAACTGCATGGTCATGGCTGAAAGTGCTAGTTGGGGGC ACATCCGTCTTTATCATTCTTGGGCTAATCGCTACAGCAGTGGTT GCCCTGGTGCTATTCACCCACAAACAC SINV mRNA Sequences SE_SINV_AR3 ORF: 57 SE_SINV_AR3 AUGGAUGUGGACGCUCACUUUACCCAGUAUAAAUUAGCCCGCCCG 39-BeAr436087_ UAUAUAGCCGACUGCCCAAAUUGCGGUCAUGGCAGAUGUGACAGU E2E1_SP CCGAUCGCUAUCGAGGACGUGCGCGGAGACGCGCACGCCGGUUAC SINV, Brazilian AUCCGCAUACAGACAUCCGCAAUGUUCGGCAUGAAGUCGGAAGGG isolate GUUGAUCUGGCUUACAUGAGCUAUAUGAACGGAAAGGUUUUAAAA GCCAUCAAGAUUGAUAGCCUGUACGUCCGUACAUCAGCACCUUGU UCUCUGGUUUCUUAUCACGGAUACUACCUCCUCGCGCAGUGCCCA CCAGGAGAUACUGUAACAGUAGGGUUCCUAGAAGGUACCCACAAG UAUAUGUGUACAGUAGCGCACCAGGUAAAAUUUAACCCGGUGGGG AGAGAGAAAUACAGACAUCCACCAGAACACGGUGUUGAACUACCC UGCAACAAAUACACCCAUAAGCGUGCCGAUCAAGGCCAUUAUGUA GAAAUGCACCAACCGGGUAUGGUCGCCGACCACACUCUGUUGAGC AUGAGCGGCACCAAGGUGAAAGUCACCGCACCGAGCAGUUCGCAA GUGAAAUAUUACUGCAAAUGUCCAGAUCUUCAAGAAGGAACUACC AGCGGUGAACACACAACAACAUGUACCGAUGUUAAGCAAUGCCGA GCGUAUCUGAUUGACAACAGGAAGUGGGUGUACAACUCAGGAAAA UUACCUAGAGGAGAAGGCGAAACCUUUAAAGGCAAACUCCAUGUA CCAUUUGUACCUGUUGCGGCCACCUGCACAGCGACCCUUGCUCCA GAGCCUCUCGUCGAGCACAAGCACCGCUCCCUGAUCCUCCACUUG CAUCCAGAGCACCCCACGCUAUUGACAACAAGGGCGCUCGGAAGU CAAGCACAACCGACUAGGCAAUGGAUAGACCGCCCAACCACCGUC AACUUCACAGUUACCGGAGAAGGUUUUGAAUAUACCUGGGGCAAC CAUCCCCCGAAAAGAGUAUGGGCCCAAGAGUCAGGAGAAGGCAAC CCGCACGGCUGGCCUCACGAGGUGGUAAUCUACUACUACAAUAGG UACCCGAUGACAACUAUCGUGGGAUUAUGCACGUGUGCCGCUAUU AUUAUGGUGUCAUGCGUUACAUCUGUAUGGCUUUUAUGCCGCACC CGUAACCUUUGCAUAACACCCUACAGAUUGGCACCAAAUGCCCAC GUACCUCUCUUAGUAGCGUUGCUGUGCUGCGUUAAACCAACUAGA GCAUACGAGCACACGGCUGUGAUGUCGAACAAGGUGGGGAUCCCU UACAAAGCCCUAGUUGAAAGGCCAGGGUACGCACCCGUACAUCUC CAGAUCCAGCUGGUAAGCACAAAAAUAAUCCCUACAGCGAACUUG GAAUACAUUACCUGUAAAUAUAAGACUAAGGUGCCUUCCCCAGUA GUAAAAUGUUGCGGAUCCACCCAAUGUACGUCUAAACAAUACCCA GACUAUCAGUGCCAAGUCUUCACGGGAGUCUACCCAUUUAUGUGG GGAGGAGCCUACUGUUUCUGUGAUACUGAAAAUACACAAAUGAGC GAAGCGUAUGUCGAACGCUCAGAAGAAUGCUCAGUGGACCAGGCC AAAGCCUACAAAGUACACACAGGAACGGUGCAGGCUGUAGUUAAU AUCACCUACGGGGGCGUCAGCUGGAGAUCUGCCGACGUCUAUGUC AACGGAGAGACCCCUGCCAAAAUAGGUGAUGCUAAACUAACUAUA GGCCCUCUCUCUUCCGCAUGGUCACCUUUUGACUCUAAAGUCAUA GUGUACGGGCACGAGGUGUAUAACUAUGACUUUCCCGAAUACGGU ACCGGCAAAGCCGGUUCGUUCGGAGACUUGCAAUCCAGGACACUU ACAAGUAAGGACUUGUACUCUAACACCAAUUUGAAACUGCAACGU CCCCAGCCAGGAGUGGUCCACACCCCAUACACCCAGGCACCCUCG GGGUUUGAACGUUGGAAGAAAGAUCGAGGGGCACCGCUAAACGAC GUCGCUCCUUUUGGAUGCAACAUAGCCCUGGAACCACUGCGUGCU GAGAACUGUGCGGUGGGGAGUAUCCCUCUCUCCAUCGACAUACCC GAUGCCGCUUUUACCAGGAUAUCGGAGACACCGACUGUCUCCGAU CUGGAAUGUAAAAUUACUGAGUGCACGUAUGCGUCAGAUUUUGGG GGAAUAGCUACUAUGUCGUAUAAAGCUAGCAAGGCAGGAAACUGC CCUAUUCAUUCCCCUUCGGGCAUUGCAGUAAUUAAAGAGAACGAU GUUAUUCUUUCUGGAAGCGGCUCGUUCACGUUCCAUUUUUCAACA GCGAGCAUCCACCCAGCGUUCAAGAUGCAGGUAUGCACCAGCGUA GUCACCUGCAAAGGUGACUGCAAGCCACCUAAGGACCAUAUCGUC GAUUACCCAGCCCAGCAUACUGAGACGUUUACAUCAGCAGUUUCC GCAACUGCAUGGUCAUGGCUGAAAGUGCUAGUUGGGGGCACAUCC GUCUUUAUCAUUCUUGGGCUAAUCGCUACAGCAGUGGUUGCCCUG GUGCUAUUCACCCACAAACAC SE_SINV_AR3 ORF: 58 SE_SINV_AR3 AUGCAGAGAGUUGUGUUUGUCGUGCUAUUGCUUUUGGUGGCCCCA 39-BeAr436087_ GCUUACAGCGAUGUGGACGCUCACUUUACCCAGUAUAAAUUAGCC E2E1 CGCCCGUAUAUAGCCGACUGCCCAAAUUGCGGUCAUGGCAGAUGU SINV, Brazilian GACAGUCCGAUCGCUAUCGAGGACGUGCGCGGAGACGCGCACGCC isolate GGUUACAUCCGCAUACAGACAUCCGCAAUGUUCGGCAUGAAGUCG GAAGGGGUUGAUCUGGCUUACAUGAGCUAUAUGAACGGAAAGGUU UUAAAAGCCAUCAAGAUUGAUAGCCUGUACGUCCGUACAUCAGCA CCUUGUUCUCUGGUUUCUUAUCACGGAUACUACCUCCUCGCGCAG UGCCCACCAGGAGAUACUGUAACAGUAGGGUUCCUAGAAGGUACC CACAAGUAUAUGUGUACAGUAGCGCACCAGGUAAAAUUUAACCCG GUGGGGAGAGAGAAAUACAGACAUCCACCAGAACACGGUGUUGAA CUACCCUGCAACAAAUACACCCAUAAGCGUGCCGAUCAAGGCCAU UAUGUAGAAAUGCACCAACCGGGUAUGGUCGCCGACCACACUCUG UUGAGCAUGAGCGGCACCAAGGUGAAAGUCACCGCACCGAGCAGU UCGCAAGUGAAAUAUUACUGCAAAUGUCCAGAUCUUCAAGAAGGA ACUACCAGCGGUGAACACACAACAACAUGUACCGAUGUUAAGCAA UGCCGAGCGUAUCUGAUUGACAACAGGAAGUGGGUGUACAACUCA GGAAAAUUACCUAGAGGAGAAGGCGAAACCUUUAAAGGCAAACUC CAUGUACCAUUUGUACCUGUUGCGGCCACCUGCACAGCGACCCUU GCUCCAGAGCCUCUCGUCGAGCACAAGCACCGCUCCCUGAUCCUC CACUUGCAUCCAGAGCACCCCACGCUAUUGACAACAAGGGCGCUC GGAAGUCAAGCACAACCGACUAGGCAAUGGAUAGACCGCCCAACC ACCGUCAACUUCACAGUUACCGGAGAAGGUUUUGAAUAUACCUGG GGCAACCAUCCCCCGAAAAGAGUAUGGGCCCAAGAGUCAGGAGAA GGCAACCCGCACGGCUGGCCUCACGAGGUGGUAAUCUACUACUAC AAUAGGUACCCGAUGACAACUAUCGUGGGAUUAUGCACGUGUGCC GCUAUUAUUAUGGUGUCAUGCGUUACAUCUGUAUGGCUUUUAUGC CGCACCCGUAACCUUUGCAUAACACCCUACAGAUUGGCACCAAAU GCCCACGUACCUCUCUUAGUAGCGUUGCUGUGCUGCGUUAAACCA ACUAGAGCAUACGAGCACACGGCUGUGAUGUCGAACAAGGUGGGG AUCCCUUACAAAGCCCUAGUUGAAAGGCCAGGGUACGCACCCGUA CAUCUCCAGAUCCAGCUGGUAAGCACAAAAAUAAUCCCUACAGCG AACUUGGAAUACAUUACCUGUAAAUAUAAGACUAAGGUGCCUUCC CCAGUAGUAAAAUGUUGCGGAUCCACCCAAUGUACGUCUAAACAA UACCCAGACUAUCAGUGCCAAGUCUUCACGGGAGUCUACCCAUUU AUGUGGGGAGGAGCCUACUGUUUCUGUGAUACUGAAAAUACACAA AUGAGCGAAGCGUAUGUCGAACGCUCAGAAGAAUGCUCAGUGGAC CAGGCCAAAGCCUACAAAGUACACACAGGAACGGUGCAGGCUGUA GUUAAUAUCACCUACGGGGGCGUCAGCUGGAGAUCUGCCGACGUC UAUGUCAACGGAGAGACCCCUGCCAAAAUAGGUGAUGCUAAACUA ACUAUAGGCCCUCUCUCUUCCGCAUGGUCACCUUUUGACUCUAAA GUCAUAGUGUACGGGCACGAGGUGUAUAACUAUGACUUUCCCGAA UACGGUACCGGCAAAGCCGGUUCGUUCGGAGACUUGCAAUCCAGG ACACUUACAAGUAAGGACUUGUACUCUAACACCAAUUUGAAACUG CAACGUCCCCAGCCAGGAGUGGUCCACACCCCAUACACCCAGGCA CCCUCGGGGUUUGAACGUUGGAAGAAAGAUCGAGGGGCACCGCUA AACGACGUCGCUCCUUUUGGAUGCAACAUAGCCCUGGAACCACUG CGUGCUGAGAACUGUGCGGUGGGGAGUAUCCCUCUCUCCAUCGAC AUACCCGAUGCCGCUUUUACCAGGAUAUCGGAGACACCGACUGUC UCCGAUCUGGAAUGUAAAAUUACUGAGUGCACGUAUGCGUCAGAU UUUGGGGGAAUAGCUACUAUGUCGUAUAAAGCUAGCAAGGCAGGA AACUGCCCUAUUCAUUCCCCUUCGGGCAUUGCAGUAAUUAAAGAG AACGAUGUUAUUCUUUCUGGAAGCGGCUCGUUCACGUUCCAUUUU UCAACAGCGAGCAUCCACCCAGCGUUCAAGAUGCAGGUAUGCACC AGCGUAGUCACCUGCAAAGGUGACUGCAAGCCACCUAAGGACCAU AUCGUCGAUUACCCAGCCCAGCAUACUGAGACGUUUACAUCAGCA GUUUCCGCAACUGCAUGGUCAUGGCUGAAAGUGCUAGUUGGGGGC ACAUCCGUCUUUAUCAUUCUUGGGCUAAUCGCUACAGCAGUGGUU GCCCUGGUGCUAUUCACCCACAAACAC

For all sequences in Table 15:

5′UTR: DNA (SEQ ID NO: 40) TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGAA ATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACC 3′UTR: DNA (SEQ ID NO: 41) TGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTC CCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCGTGGTCTTTGAA TAAAGTCTGAGTGGGCGGC 5′UTR: mRNA (SEQ ID NO: 42) UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGGGAA AUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACC 3′UTR: mRNA (SEQ ID NO: 43) UGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUC CCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCGUGGUCUUUGAA UAAAGUCUGAGUGGGCGGC

TABLE 16 SINV Amino Acid Sequences Antigen Amino Acid Sequence SE_SINV_AR3 MDVDAHFTQYKLARPYIADCPNCGHGRCDSPIAIEDVRGDAHAGYI 59 39-BeAr436087_ RIQTSAMFGMKSEGVDLAYMSYMNGKVLKAIKIDSLYVRTSAPCSL E2E1_SP VSYHGYYLLAQCPPGDTVTVGFLEGTHKYMCTVAHQVKFNPVGREK YRHPPEHGVELPCNKYTHKRADQGHYVEMHQPGMVADHTLLSMSGT KVKVTAPSSSQVKYYCKCPDLQEGTTSGEHTTTCTDVKQCRAYLID NRKWVYNSGKLPRGEGETFKGKLHVPFVPVAATCTATLAPEPLVEH KHRSLILHLHPEHPTLLTTRALGSQAQPTRQWIDRPTTVNFTVTGE GFEYTWGNHPPKRVWAQESGEGNPHGWPHEVVIYYYNRYPMTTIVG LCTCAAIIMVSCVTSINVWLLCRTRNLCITPYRLAPNAHVPLLVAL LCCVKPTRAYEHTAVMSNKVGIPYKALVERPGYAPVHLQIQLVSTK IIPTANLEYITCKYKTKVPSPVVKCCGSTQCTSKQYPDYQCQVFTG VYPFMWGGAYCFCDTENTQMSEAYVERSEECSINVDQAKAYKVHTG TVQAVVNITYGGVSWRSADVYVNGETPAKIGDAKLTIGPLSSAWSP FDSKVIVYGHEVYNYDFPEYGTGKAGSFGDLQSRTLTSKDLYSNTN LKLQRPQPGVVHTPYTQAPSGFERWKKDRGAPLNDVAPFGCNIALE PLRAENCAVGSIPLSIDIPDAAFTRISETPTVSDLECKITECTYAS DFGGIATMSYKASKAGNCPIHSPSGIAVIKENDVILSGSGSFTFHF STASIHPAFKMQVCTSINVVTCKGDCKPPKDHIVDYPAQHTETFTS AVSATAWSWLKVLVGGTSINVFIILGLIATAVVALVLFTHKH SE_SINV_AR3 MQRVVFVVLLLLVAPAYSDVDAHFTQYKLARPYIADCPNCGHGRCD 60 39-BeAr436087_ SPIAIEDVRGDAHAGYIRIQTSAMFGMKSEGVDLAYMSYMNGKVLK E2E1 AIKIDSLYVRTSAPCSLVSYHGYYLLAQCPPGDTVTVGFLEGTHKY MCTVAHQVKFNPVGREKYRHPPEHGVELPCNKYTHKRADQGHYVEM HQPGMVADHTLLSMSGTKVKVTAPSSSQVKYYCKCPDLQEGTTSGE HTTTCTDVKQCRAYLIDNRKWVYNSGKLPRGEGETFKGKLHVPFVP VAATCTATLAPEPLVEHKHRSLILHLHPEHPTLLTTRALGSQAQPT RQWIDRPTTVNFTVTGEGFEYTWGNHPPKRVWAQESGEGNPHGWPH EVVIYYYNRYPMTTIVGLCTCAAIIMVSCVTSINVWLLCRTRNLCI TPYRLAPNAHVPLLVALLCCVKPTRAYEHTAVMSNKVGIPYKALVE RPGYAPVHLQIQLVSTKIIPTANLEYITCKYKTKVPSPVVKCCGST QCTSKQYPDYQCQVFTGVYPFMWGGAYCFCDTENTQMSEAYVERSE ECSINVDQAKAYKVHTGTVQAVVNITYGGVSWRSADVYVNGETPAK IGDAKLTIGPLSSAWSPFDSKVIVYGHEVYNYDFPEYGTGKAGSFG DLQSRTLTSKDLYSNTNLKLQRPQPGVVHTPYTQAPSGFERWKKDR GAPLNDVAPFGCNIALEPLRAENCAVGSIPLSIDIPDAAFTRISET PTVSDLECKITECTYASDFGGIATMSYKASKAGNCPIHSPSGIAVI KENDVILSGSGSFTFHFSTASIHPAFKMQVCTSINVVTCKGDCKPP KDHIVDYPAQHTETFTSAVSATAWSWLKVLVGGTSINVFIILGLIA TAVVALVLFTHKH SE_SINV_E2 KPPSGKNITYECKCGDYKTATVSINVRTEIAGCTAIKQCVAYKSDQ 61 AF439980_1 TKWVFNSPDLIRHADHTAQGKLHLPFKPTLSTCLVPLAHEPTVTHG polyprotein FKHISLHLDTDHPTLLTTRRLGEKPEPTSEWISGKTVRNFTVDRDG [Sindbis virus] L SE_SINV_AR3 MQRVVFVVLLLLVAPAYSDVDAHFTQYKLARPYIADCPNCGHGRCD 62 39-BeAr436087_ SPIAIEDVRGDAHAGYIRIQTSAMFGMKSEGVDLAYMSYMNGKVLK E2E1 AIKIDSLYVRTSAPCSLVSYHGYYLLAQCPPGDTVTVGFLEGTHKY MCTVAHQVKFNPVGREKYRHPPEHGVELPCNKYTHKRADQGHYVEM HQPGMVADHTLLSMSGTKVKVTAPSSSQVKYYCKCPDLQEGTTSGE HTTTCTDVKQCRAYLIDNRKWVYNSGKLPRGEGETFKGKLHVPFVP VAATCTATLAPEPLVEHKHRSLILHLHPEHPTLLTTRALGSQAQPT RQWIDRPTTVNFTVTGEGFEYTWGNHPPKRVWAQESGEGNPHGWPH EVVIYYYNRYPMTTIVGLCTCAAIIMVSCVTSINVWLLCRTRNLCI TPYRLAPNAHVPLLVALLCCVKPTRAYEHTAVMSNKVGIPYKALVE RPGYAPVHLQIQLVSTKIIPTANLEYITCKYKTKVPSPVVKCCGST QCTSKQYPDYQCQVFTGVYPFMWGGAYCFCDTENTQMSEAYVERSE ECSINVDQAKAYKVHTGTVQAVVNITYGGVSWRSADVYVNGETPAK IGDAKLTIGPLSSAWSPFDSKVIVYGHEVYNYDFPEYGTGKAGSFG DLQSRTLTSKDLYSNTNLKLQRPQPGVVHTPYTQAPSGFERWKKDR GAPLNDVAPFGCNIALEPLRAENCAVGSIPLSIDIPDAAFTRISET PTVSDLECKITECTYASDFGGIATMSYKASKAGNCPIHSPSGIAVI KENDVILSGSGSFTFHFSTASIHPAFKMQVCTSINVVTCKGDCKPP KDHIVDYPAQHTETFTSAVSATAWSWLKVLVGGTSINVFIILGLIA TAVVALVLFTHKH SE_SINV_AR3 MDVDAHFTQYKLARPYIADCPNCGHGRCDSPIAIEDVRGDAHAGYI 63 39-BeAr436087_ RIQTSAMFGMKSEGVDLAYMSYMNGKVLKAIKIDSLYVRTSAPCSL E2E1_SP VSYHGYYLLAQCPPGDTVTVGIFLEGTHKYMCTVAHQVKFNPVGRE KYRHPPEHGVELPCNKYTHKRADQGHYVEMHQPGMVADHTLLSMSG TKVKVTAPSSSQVKYYCKCPDLQEGTTSGEHTTTCTDVKQCRAYLI DNRKWVYNSGKLPRGEGETFKGKLHVPFVPVAATCTATLAPEPLVE HKHRSLILHLHPEHPTLLTTRALGSQAQPTRQWIDRPTTVNFTVTG EGFEYTWGNHPPKRVWAQESGEGNPHGWPHEVVIYYYNRYPMTTIV GLCTCAAIIMVSCVTSINVWLLCRTRNLCITPYRLAPNAHVPLLVA LLCCVKPTRAYEHTAVMSNKVGIPYKALVERPGYAPVHLQIQLVST KIIPTANLEYITCKYKTKVPSPVVKCCGSTQCTSKQYPDYQCQVFT GVYPFMWGGAYCFCDTENTQMSEAYVERSEECSINVDQAKAYKVHT GTVQAVVNITYGGVSWRSADVYVNGETPAKIGDAKLTIGPLSSAWS PFDSKVIVYGHEVYNYDFPEYGTGKAGSFGDLQSRTLTSKDLYSNT NLKLQRPQPGVVHTPYTQAPSGFERWKKDRGAPLNDVAPFGCNIAL EPLRAENCAVGSIPLSIDIPDAAFTRISETPTVSDLECKITECTYA SDFGGIATMSYKASKAGNCPIHSPSGIAVIKENDVILSGSGSFTFH FSTASIHPAFKMQVCTSINVVTCKGDCKPPKDHIVDYPAQHTETFT SAVSATAWSWLKVLVGGTSINVFIILGLIATAVVALVLFTHKH

TABLE 17 SINV strains/isolates, structural proteins/variants - Homo sapiens GenBank Virus Strain Accession E2 structural protein [Sindbis virus] AAB07099.1 E2 structural protein [Sindbis virus] AAB07111.1 E2 structural protein [Sindbis virus] AAB07084.1 E2 structural protein [Sindbis virus] AAB07080.1 E2 protein [Sindbis virus] AIS25577.1 E2 structural protein [Sindbis virus] AAB07093.1 E2 structural protein [Sindbis virus] AAB07102.1 E2 structural protein [Sindbis virus] AAB07103.1 E2 protein [Sindbis virus] AIS25580.1 E2 protein [Sindbis virus] AGI50355.1 E2 structural protein [Sindbis virus] AAB07090.1 E2 structural protein [Sindbis virus] AAB07097.1 E2 structural protein [Sindbis virus] AAB07104.1 E2 structural protein [Sindbis virus] AAB07110.1 E2 structural protein [Sindbis virus] AAB07098.1 E2 structural protein [Sindbis virus] AAB07078.1 E2 protein [Sindbis virus] AGI50359.1 E2 structural protein [Sindbis virus] AAB07096.1 E2 structural protein [Sindbis virus] AAB07082.1 E2 structural protein [Sindbis virus] AAB07105.1 E2 structural protein [Sindbis virus] AAB07092.1 E2 structural protein [Sindbis virus] AAB07101.1 E2 structural protein [Sindbis virus] AAB07109.1 E2 structural protein [Sindbis virus] AAB07085.1 E2 structural protein [Sindbis virus] AAB07075.1 E2 structural protein [Sindbis virus] AAB07086.1 E2 structural protein [Sindbis virus] AAB07081.1 structural polyprotein [Sindbis virus] AAA47485.1 E2 structural protein [Sindbis virus] AAB07089.1 E2 structural protein [Sindbis virus] AAB07106.1 Chain U, Pseudo-Atomic Structure Of The 3MUW_U E2-E1 Protein Shell In Sindbis Virus E2 structural protein [Sindbis virus] AAB07094.1 E2 structural protein [Sindbis virus] AAB07095.1 E2 structural protein [Sindbis virus] AAB07107.1 E2 structural protein [Sindbis virus] AAB07087.1 e-2 structural protein [Sindbis virus] NP_740675.1 structural polyprotein [Sindbis virus] ACU25469.1 glycoprotein PE2 [Sindbis virus] AAA79886.1 structural polyprotein [Sindbis virus] ADC34086.1 structural polyprotein [Sindbis virus] ADC34084.1 structural polyprotein [Sindbis virus] AFL65790.1 structural polyprotein [Sindbis-like virus] AAA86136.1 structural polyprotein [Sindbis virus] AHC94941.1 Structural polyprotein; Contains: Capsid P27285.1 protein; Coat protein; p62; E3/E2; E3 protein; Spike glycoprotein E3; E2 envelope glycoprotein; 6K protein; E1 envelope glycoprotein structural polyprotein [Sindbis-like virus] AAA86134.1 structural polyprotein [Sindbis virus] AGN55417.1 structural polyprotein [Sindbis virus] AFL65800.1 structural polyprotein [Sindbis virus] AFL65796.1 structural polyprotein [Sindbis virus] AFL65794.1 structural polyprotein [Sindbis virus] AFL65792.1 structural polyprotein [Sindbis virus] AFL65788.1 structural polyprotein [Sindbis virus] ACU25466.1 structural polyprotein precursor [Babanki virus] AAO33325.1 structural polyprotein [Babanki virus] AEJ36222.1 structural polyprotein [Sindbis virus] AFL65798.1 p130 structural polyprotein [Sindbis-like virus YN87448] AAC77466.1 structural polyprotein [Sindbis virus] AKZ17416.1 structural polyprotein [Sindbis virus] AKZ17540.1 structural polyprotein [Sindbis virus] AKZ17497.1 structural polyprotein [Sindbis virus] AKZ17377.1 structural polyprotein [Sindbis virus] AKZ17356.1 structural polyprotein [Sindbis virus] AKZ17290.1 structural polyprotein [Sindbis virus] AKZ17211.1 structural polyprotein [Sindbis virus] AKZ17371.1 structural polyprotein [Sindbis virus] AKZ17344.1 structural polyprotein [Sindbis virus] AKZ17332.1 structural polyprotein [Sindbis virus] AKZ17464.1 structural polyprotein [Sindbis virus] AKZ17235.1 structural polyprotein [Sindbis virus] AKZ17202.1 structural polyprotein [Sindbis virus] AKZ17187.1 structural polyprotein [Sindbis virus] AKZ17178.1 structural polyprotein [Sindbis virus] AKZ17558.1 structural polyprotein [Sindbis virus] AKZ17506.1 structural polyprotein [Sindbis virus] AKZ17269.1 structural polyprotein [Sindbis virus] AKZ17482.1 structural polyprotein [Sindbis virus] AKZ17467.1 structural polyprotein [Sindbis virus] AKZ17368.1 structural polyprotein [Sindbis virus] AKZ17359.1 structural polyprotein [Sindbis virus] AKZ17347.1 structural polyprotein [Sindbis virus] AKZ17341.1 structural polyprotein [Sindbis virus] AKZ17470.1 structural polyprotein [Sindbis virus] AKZ17528.1 structural polyprotein [Sindbis virus] AKZ17522.1 structural polyprotein [Sindbis virus] AKZ17386.1 structural polyprotein [Sindbis virus] AKZ17223.1 structural polyprotein [Sindbis virus] AKZ17594.1 structural polyprotein [Sindbis virus] AKZ17317.1 structural polyprotein [Sindbis virus] AKZ17196.1 structural polyprotein [Sindbis virus] AKZ17281.1 structural polyprotein [Sindbis virus] AKZ17479.1 structural polyprotein [Sindbis virus] AKZ17452.1

TABLE 18 VEEV Amino Acid Sequence Antigen Amino Acid Sequence SEQ ID NO: Venezuelan equine MFPFQPMYPMQPMPYRNPFAAPRRPWFPRTDPFLAMQVQELTRS 64 encephalitis virus MANLTFKQRRDAPPEGPSAKKPKKEASQKQKGGGQGKKKKNQGK Structural poly KKAKTGPPNPKAQNGNKKKTNKKPGKRQRMVMKLESDKTFPIML protein product = EGKINGYACVVGGKLFRPMHVEGKIDNDVLAALKTKKASKYDLE “structural YADVPQNMRADTFKYTHEKPQGYYSWHHGAVQYENGRFTVPKGV polyprotein” GAKGDSGRPILDNQGRVVAIVLGGVNEGSRTALSINVVMWNEKG /protein_id = VTVKYTPENCEQWSLVTTMCLLANVTFPCAQPPICYDRKPAETL “AAC19322.1” AMLSINVNVDNPGYDELLEAAVKCPGRKRRSTEELFKEYKLTRP /db_xref = YMARCIRCAVGSCHSPIAIEAVKSDGHDGYVRLQTSSQYGLDSS “GI: 1375090” GNLKGRTMRYDMHGTIKEIPLHQVSLHTSRPCHIVDGHGYFLLA RCPAGDSITMEFKKADSINVTHSCSINVPYEVKFNPVGRELYTH PPEHGVEQACQVYAHDAQNRGAYVEMHLPGSEVDSSLVSLSGSS INVTVTPPVGTSALVECECGGTKISETINKTKQFSQCTKKEQCR AYRLQNDKWVYNSDKLPKAAGATLKGKLHVPFLLADGKCTVPLA PEPMITFGFRSINVSLKLHPKNPTYLTTRQLADEPHYTHELISE PAVRNFTVTEKGWEFVWGNHPPKRFWAQETAPGNPHGLPHEVIT HYYHRYPMSTILGLSICAAIATVSINVAASTWLFCRSRVACLTP YRLTPNARIPFCLAVLCCARTARAETTWESLDHLWNNNQQMFWI QLLIPLAALIVVTRLLRCVCCVVPFLVMAGAAGAGAYEHATTMP SQAGISYNTIVNRAGYAPLPISITPTKIKLIPTVNLEYVTCHYK TGMDSPAIKCCGSQECTPTYRPDEQCKVFTGVYPFMWGGAYCFC DTENTQVSKAYVMKSDDCLADHAEAYKAHTASINVQAFLNITVG EHSIVTTVYVNGETPVNFNGVKLTAGPLSTAWTPFDRKIVQYAG EIYNYDFPEYGAGQPGAFGDIQSRTVSSSDLYANTNLVLQRPKA GAIHVPYTQAPSGFEQWKKDKAPSLKFTAPFGCEIYTNPIRAEN CAVGSIPLAFDIPDALFTRVSETPTLSAAECTLNECVYSSDFGG IATVKYSASKSGKCAVHVPSGTATLKEAAVELTEQGSATIHFST ANIHPEFRLQICTSYVTCKGDCHPPKDHIVTHPQYHAQTFTAAV SKTAWTWLTSLLGGSAVIIIIGLVLATIVAMYVLTNQKHN

TABLE 19 Example 19 Test Conditions BALB/C Immunization Group Vaccine n Dose Route Prime Boost Endpoint 1 Yellow fever 5 10 ug IM Day 0 Day 21 Terminal bleeds on prME vaccine Day 41. Anti 2 Yellow fever 5 10 ug IM Day 0 NA Yellow fever prME vaccine neutralizing IgG titer. 3 Yellow fever 5  2 ug IM Day 0 Day 21 prME vaccine 4 Yellow fever 5  2 ug IM Day 0 NA prME vaccine 5 PBS 5 NA IM Day 0 Day 21

TABLE 20 Example 20 Test Conditions AG129 Immunization Group Vaccine n Dose Route Prime Boost Challenge Endpoint 1 Yellow fever 8 10 ug IM Day 0 Day 21 Day 42 Monitor for prME vaccine survival 2 Yellow fever 8 10 ug IM Day 0 NA and prME vaccine weightloss. 3 Yellow fever 8  2 ug IM Day 0 Day 21 Viral load prME vaccine at Day 5 4 Yellow fever 8  2 ug IM Day 0 NA prME vaccine 5 PBS 8 NA IM Day 0 Day 21

TABLE 21 YFV Nucleic Acid Sequences Description Sequence SEQ ID NO: YF_Con_prME_ ATGGAAACCCCTGCCCAGCTGCTGTTCCTGCTGCTGCTGTGGCTGC 65 Sen2000_IgGksp CTGATACCACCGGCGTGACACTCGTGCGGAAGAACAGATGGCTGCT GCTGAACGTGACCAGCGAGGACCTGGGCAAGACCTTCTCTGTGGGC ACCGGCAACTGCACCACCAACATCCTGGAAGCCAAGTACTGGTGCC CCGACAGCATGGAGTACAACTGCCCCAACCTGAGCCCCAGAGAGGA ACCCGACGACATCGACTGCTGGTGCTACGGCGTGGAAAACGTGCGG GTGGCCACGGCAAGTGCGATAGCGCCGGCAGAAGCAGAAGAAGCAG GCGGGCCATCGACCTGCCCACCCACGAAAACCACGGCCTGAAAACC CGGCAGGAAAAGTGGATGACCGGCCGGATGGGCGAGCGGCAGCTGC AGAAAATCGAGAGATGGCTCGTGCGCAACCCCTTCTTCGCCGTGAC CGCCCTGACAATCGCCTACCTCGTGGGCAGCAACATGACCCAGAGA GTCGTGATCGCCCTGCTGGTGCTGGCTGTGGGCCCTGCCTATAGCG CCCACTGTATCGGCATCACCGACCGGGACTTCATCGAGGGCGTGCA CGGCGGAACATGGGTGTCCGCTACCCTGGAACAGGATAAGTGCGTG ACCGTGATGGCCCCCGACAAGCCCAGCCTGGACATCAGCCTGGAAA CCGTGGCCATTGACGGACCCGCCGAGGCCAGAAAAGTGTGCTACAA CGCCGTGCTGACCCACGTGAAGATCAACGACAAGTGCCCCAGCACC GGCGAAGCCCACCTGGCCGAAGAGAACGAGGGCGACAACGCCTGCA AGCGGACCTACAGCGATAGAGGCTGGGGCAATGGCTGCGGCCTGTT TGGCAAGGGCAGCATCGTGGCCTGCGCCAAGTTCACCTGTGCCAAG AGCATGAGCCTGTTCGAGGTGGACCAGACCAAGATCCAGTACGTGA TCCGGGCTCAGCTGCACGTGGGCGCCAAGCAGGAAAACTGGAACAC CGACATCAAGACCCTGAAGTTCGACGCCCTGAGCGGCTCCCAGGAA GCCGAGTTTACCGGCTATGGCAAGGCCACCCTGGAATGCCAGGTGC AGACCGCCGTGGACTTCGGCAACAGCTATATCGCCGAGATGGAAAA AGAAAGCTGGATCGTGGACCGGCAGTGGGCCCAGGATCTGACACTG CCTTGGCAGTCTGGCTCTGGCGGAGTGTGGCGGGAAATGCACCACC TGGTGGAATTCGAGCCTCCCCACGCCGCCACCATTAGAGTGCTGGC CCTGGGCAATCAGGAAGGCTCTCTGAAAACAGCCCTGACCGGCGCC ATGAGAGTGACCAAGGACACCAACGACAACAACCTGTACAAGCTGC ACGGGGGGCACGTGTCCTGCAGAGTGAAACTGTCTGCCCTGACACT GAAGGGCACCAGCTACAAGATGTGCACCGACAAGATGAGCTTCGTG AAGAACCCCACCGACACCGGCCACGGCACAGTCGTGATGCAAGTGA AGGTGCCCAAGGGCGCTCCCTGCAGAATCCCTGTGATCGTGGCCGA TGATCTGACAGCCGCCATCAACAAGGGCATCCTCGTGACAGTGAAC CCTATCGCCTCCACCAACGATGACGAGGTGCTGATCGAAGTGAACC CCCCCTTCGGCGACTCCTACATCATCGTGGGCACAGGCGACAGCAG ACTGACCTACCAGTGGCACAAAGAGGGCAGCAGCATCGGCAAGCTG TTCACCCAGACCATGAAGGGCGCCGAGAGACTGGCTGTGATGGGAG ATGCCGCCTGGGACTTTAGCAGCGCTGGCGGCTTCTTTACCAGCGT GGGCAAGGGAATCCACACCGTGTTCGGCAGCGCCTTCCAGGGACTG TTTGGCGGCCTGAGCTGGATCACCAAAGTGATCATGGGCGCTGTGC TGATCTGGGTGGGAATCAACACCCGGAACATGACCATGAGCATGTC CATGATCCTCGTGGGAGTGATTATGATGTTCCTGAGCCTGGGCGTG GGAGCC YF_Con_prME_ ATGTGGCTGGTGTCCCTGGCCATCGTGACAGCCTGTGCTGGCGCTG 66 Sen2000_JEVsp TGACACTCGTGCGGAAGAACAGATGGCTGCTGCTGAACGTGACCAG CGAGGACCTGGGCAAGACCTTCTCTGTGGGCACCGGCAACTGCACC ACCAACATCCTGGAAGCCAAGTACTGGTGCCCCGACAGCATGGAGT ACAACTGCCCCAACCTGAGCCCCAGAGAGGAACCCGACGACATCGA CTGCTGGTGCTACGGCGTGGAAAACGTGCGGGTGGCCTACGGCAAG TGCGATAGCGCCGGCAGAAGCAGAAGAAGCAGGCGGGCCATCGACC TGCCCACCCACGAAAACCACGGCCTGAAAACCCGGCAGGAAAAGTG GATGACCGGCCGGATGGGCGAGCGGCAGCTGCAGAAAATCGAGAGA TGGCTCGTGCGCAACCCCTTCTTCGCCGTGACCGCCCTGACAATCG CCTACCTCGTGGGCAGCAACATGACCCAGAGAGTCGTGATCGCCCT GCTGGTGCTGGCTGTGGGCCCTGCCTATAGCGCCCACTGTATCGGC ATCACCGACCGGGACTTCATCGAGGGCGTGCACGGCGGAACATGGG TGTCCGCTACCCTGGAACAGGATAAGTGCGTGACCGTGATGGCCCC CGACAAGCCCAGCCTGGACATCAGCCTGGAAACCGTGGCCATTGAC GGACCCGCCGAGGCCAGAAAAGTGTGCTACAACGCCGTGCTGACCC ACGTGAAGATCAACGACAAGTGCCCCAGCACCGGCGAAGCCCACCT GGCCGAAGAGAACGAGGGCGACAACGCCTGCAAGCGGACCTACAGC GATAGAGGCTGGGGCAATGGCTGCGGCCTGTTTGGCAAGGGCAGAC ATCGTGGCCTGCGCCAAGTTCACCTGTGCCAAGAGCATGAGCCTGT TCGAGGTGGACCAGACCAAGATCCAGTACGTGATCCGGGCCCAGCT GCACGTGGGAGCCAAGCAGGAAAACTGGAACACCGACATCAAGACC CTGAAGTTCGACGCCCTGAGCGGCTCCCAGGAAGCCGAGTTTACCG GCTATGGCAAGGCCACCCTGGAATGCCAGGTGCAGACCGCCGTGGA CTTCGGCAACAGCTATATCGCCGAGATGGAAAAAGAAAGCTGGATC GTGGACCGGCAGTGGGCCCAGGACCTGACACTGCCTTGGCAGTCTG GATCTGGCGGCGTGTGGCGGGAAATGCACCACCTGGTGGAATTCGA GCCTCCCCATGCCGCCACCATCAGAGTGCTGGCCCTGGGCAATCAG GAAGGCTCTCTGAAAACAGCCCTGACCGGCGCCATGAGAGTGACCA AGGACACCAACGACAACAACCTGTACAAGCTGCATGGCGGCCACGT GTCCTGCAGAGTGAAGCTGTCTGCCCTGACCCTGAAAGGCACCAGC TACAAGATGTGCACCGACAAGATGAGCTTCGTGAAGAACCCCACCG ACACCGGCCACGGCACAGTCGTGATGCAAGTGAAGGTGCCCAAGGG CGCTCCCTGCAGAATCCCTGTGATCGTGGCCGATGATCTGACAGCC GCCATCAACAAGGGCATCCTCGTGACAGTGAACCCTATCGCCTCCA CCAACGATGACGAGGTGCTGATCGAAGTGAACCCCCCCTTCGGCGA CTCCTACATCATCGTGGGCACAGGCGACAGCAGACTGACCTACCAG TGGCACAAAGAGGGCAGCAGCATCGGCAAGCTGTTCACCCAGACCA TGAAGGGCGCCGAGAGACTGGCTGTGATGGGAGATGCCGCCTGGGA CTTTAGCAGCGCTGGCGGCTTCTTTACCAGCGTGGGCAAGGGAATC CACACCGTGTTCGGCAGCGCCTTCCAGGGACTGTTTGGCGGCCTGA GCTGGATCACCAAAGTGATCATGGGCGCTGTGCTGATCTGGGTGGG AATCAACACCCGGAACATGACCATGAGCATGTCCATGATCCTCGTG GGAGTGATTATGATGTTCCTGAGCCTGGGCGTGGGCGCC YF_Con_prME_ ATGTGGCTGGTGTCCCTGGCCATCGTGACAGCCTGTGCTGGCGCTG 67 Sen2000_JEVsp_ TGACACTCGTGCGGAAGAACAGATGGCTGCTGCTGAACGTGACCAG N153T CGAGGACCTGGGCAAGACCTTCTCTGTGGGCACCGGCAACTGCACC ACCAACATCCTGGAAGCCAAGTACTGGTGCCCCGACAGCATGGAGT ACAACTGCCCCAACCTGAGCCCCAGAGAGGAACCCGACGACATCGA CTGCTGGTGCTACGGCGTGGAAAACGTGCGGGTGGCCTACGGCAAG TGCGATAGCGCCGGCAGAAGCAGAAGAAGCAGGCGGGCCATCGACC TGCCCACCCACGAAAACCACGGCCTGAAAACCCGGCAGGAAAAGTG GATGACCGGCCGGATGGGCGAGCGGCAGCTGCAGAAAATCGAGAGA TGGCTCGTGCGCAACCCCTTCTTCGCCGTGACCGCCCTGACAATCG CCTACCTCGTGGGCAGCAACATGACCCAGAGAGTCGTGATCGCCCT GCTGGTGCTGGCTGTGGGCCCTGCCTATAGCGCCCACTGTATCGGC ATCACCGACCGGGACTTCATCGAGGGCGTGCACGGCGGAACATGGG TGTCCGCTACCCTGGAACAGGATAAGTGCGTGACCGTGATGGCCCC CGACAAGCCCAGCCTGGACATCAGCCTGGAAACCGTGGCCATTGAC GGACCCGCCGAGGCCAGAAAAGTGTGCTACAACGCCGTGCTGACCC ACGTGAAGATCAACGACAAGTGCCCCAGCACCGGCGAAGCCCACCT GGCCGAAGAGAACGAGGGCGACAACGCCTGCAAGCGGACCTACAGC GATAGAGGCTGGGGCAATGGCTGCGGCCTGTTTGGCAAGGGCAGCA TCGTGGCCTGCGCCAAGTTCACCTGTGCCAAGAGCATGAGCCTGTT CGAGGTGGACCAGACCAAGATCCAGTACGTGATCCGGGCCCAGCTG CACGTGGGAGCCAAGCAGGAAAACTGGACCACCGACATCAAGACCC TGAAGTTCGACGCCCTGAGCGGCTCCCAGGAAGCCGAGTTTACCGG CTATGGCAAGGCCACCCTGGAATGCCAGGTGCAGACCGCCGTGGAC TTCGGCAACAGCTATATCGCCGAGATGGAAAAAGAAAGCTGGATCG TGGACCGGCAGTGGGCCCAGGACCTGACACTGCCTTGGCAGTCTGG ATCTGGCGGCGTGTGGCGGGAAATGCACCACCTGGTGGAATTCGAG CCTCCCCATGCCGCCACCATCAGAGTGCTGGCCCTGGGCAATCAGG AAGGCTCTCTGAAAACAGCCCTGACCGGCGCCATGAGAGTGACCAA GGACACCAACGACAACAACCTGTACAAGCTGCATGGCGGCCACGTG TCCTGCAGAGTGAAGCTGTCTGCCCTGACCCTGAAAGGCACCAGCT ACAAGATGTGCACCGACAAGATGAGCTTCGTGAAGAACCCCACCGA CACCGGCCACGGCACAGTCGTGATGCAAGTGAAGGTGCCCAAGGGC GCTCCCTGCAGAATCCCTGTGATCGTGGCCGATGATCTGACAGCCG CCATCAACAAGGGCATCCTCGTGACAGTGAACCCTATCGCCTCCAC CAACGATGACGAGGTGCTGATCGAAGTGAACCCCCCCTTCGGCGAC TCCTACATCATCGTGGGCACAGGCGACAGCAGACTGACCTACCAGT GGCACAAAGAGGGCAGCAGCATCGGCAAGCTGTTCACCCAGACCAT GAAGGGCGCCGAGAGACTGGCTGTGATGGGAGATGCCGCCTGGGAC TTTAGCAGCGCTGGCGGCTTCTTTACCAGCGTGGGCAAGGGAATCC ACACCGTGTTCGGCAGCGCCTTCCAGGGACTGTTTGGCGGCCTGAG CTGGATCACCAAAGTGATCATGGGCGCTGTGCTGATCTGGGTGGGA ATCAACACCCGGAACATGACCATGAGCATGTCCATGATCCTCGTGG GAGTGATTATGATGTTCCTGAGCCTGGGCGTGGGCGCC 17D_vaccine_ ATGTGGCTGGTGTCCCTGGCCATCGTGACAGCCTGTGCTGGCGCTG 68 prME_JEVsp TGACACTCGTGCGGAAAAACAGATGGCTGCTGCTGAACGTGACCAG CGAGGACCTGGGCAAGACCTTCTCTGTAGGCACCGGCAACTGCACC ACCAACATCCTGGAAGCCAAGTACTGGTGCCCCGACAGCATGGAGT ACAACTGCCCCAACCTGAGCCCCAGAGAGGAACCCGACGACATCGA CTGCTGGTGCTACGGCGTGGAAAACGTGCGGGTGGCCTACGGCAAG TGCGATAGCGCCGGCAGAAGCAGAAGAAGCAGGCGGGCCATCGACC TGCCCACCCACGAAAACCACGGCCTGAAAACCCGGCAGGAAAAGTG GATGACCGGCCGGATGGGCGAGCGGCAGCTGCAGAAAATTGAGCGG TGGTTTGTGCGGAACCCCTTCTTCGCCGTGACCGCCCTGACAATCG CCTACCTCGTGGGCAGCAACATGACCCAGAGAGTCGTGATCGCCCT GCTGGTGCTGGCTGTGGGCCCTGCCTATAGCGCCATTGATCTGCCT ACACACGAGAATCATGGGCTGAAAACAAGACAGGAAAAATGGATGA CTGGGCGCATGGGAGAAAGACAGCTGCAGAAAATCGAACGGTGGTT CGTGCGCAATCCTTTTTTTGCTGTGACTGCTCTGACCATTGCCTAT CTCGTGGGATCCAATATGACACAGCGGGTCGTGATTGCTCTGCTGG TGCTGGCAGTGGGACCCGCTTACTCCGCCCACTGTATCGGCATCAC CGACCGGGACTTCATCGAGGGCGTGCACGGCGGAACATGGGTGTCC GCTACCCTGGAACAGGATAAGTGCGTGACCGTGATGGCCCCCGACA AGCCCAGCCTGGACATCAGCCTGGAAACCGTGGCCATCGATAGACC CGCCGAAGTGCGGAAAGTGTGCTACAACGCCGTGCTGACCCACGTG AAGATCAACGACAAGTGCCCCAGCACCGGCGAAGCCCACCTGGCCG AAGAGAACGAGGGCGACAACGCCTGCAAGCGGACCTACAGCGATAG AGGCTGGGGCAATGGCTGCGGCCTGTTTGGCAAGGGCAGCATCGTG GCCTGCGCCAAGTTCACCTGTGCCAAGAGCATGAGCCTGTTCGAGG TGGACCAGACCAAGATCCAGTACGTGATCCGGGCCCAGCTGCACGT GGGAGCCAAGCAGGAAAACTGGAACACCGACATCAAGACCCTGAAG TTCGACGCCCTGAGCGGCTCCCAGGAAGTGGAATTCATCGGCTATG GCAAGGCCACCCTGGAATGTCAGGTGCAGACCGCCGTGGACTTCGG CAACAGCTATATCGCCGAGATGGAAACCGAGAGCTGGATCGTGGAC CGGCAGTGGGCTCAGGATCTGACCCTGCCTTGGCAGTCTGGCTCTG GCGGAGTGTGGCGGGAAATGCACCACCTGGTGGAATTCGAGCCTCC CCACGCCGCCACCATTAGAGTGCTGGCCCTGGGCAATCAGGAAGGC TCTCTGAAAACAGCCCTGACCGGCGCCATGAGAGTGACCAAGGACA CCAACGACAACAACCTGTACAAGCTGCATGGCGGCCACGTGTCCTG CAGAGTGAAGCTGTCTGCCCTGACACTGAAGGGCACCAGCTACAAG ATCTGCACCGACAAGATGTTCTTCGTGAAGAACCCCACCGACACCG GCCACGGCACAGTCGTGATGCAAGTGAAGGTGTCCAAGGGCGCTCC CTGCCGGATCCCTGTGATCGTGGCCGATGATCTGACAGCCGCCATC AACAAGGGCATCCTCGTGACAGTGAACCCTATCGCCTCCACCAACG ATGACGAGGTGCTGATCGAAGTGAACCCCCCCTTCGGCGACTCCTA CATCATCGTGGGACGGGGCGACAGCAGACTGACCTACCAGTGGCAC AAAGAGGGCAGCAGCATCGGCAAGCTGTTCACCCAGACCATGAAGG GCGTGGAACGGCTGGCCGTGATGGGAGATACCGCCTGGGATTTCTC TAGCGCTGGCGGCTTCTTCACCAGCGTGGGCAAGGGAATCCACACC GTGTTCGGCAGCGCCTTCCAGGGACTGTTCGGCGGCCTGAACTGGA TCACCAAAGTGATCATGGGCGCTGTGCTGATCTGGGTGGGAATCAA CACCCGGAACATGACCATGAGCATGTCCATGATCCTCGTGGGAGTG ATTATGATGTTCCTGAGCCTGGGTGTGGGCGCC 17D_vaccine_ ATGGAAACCCCTGCCCAGCTGCTGTTCCTGCTGCTGCTGTGGCTGC 69 prME_IgGKsp CTGATACCACCGGCGTGACACTCGTGCGGAAGAACAGATGGCTGCT GCTGAACGTGACCAGCGAGGACCTGGGCAAGACCTTCTCTGTGGGC ACCGGCAACTGCACCACCAACATCCTGGAAGCCAAGTACTGGTGCC CCGACAGCATGGAGTACAACTGCCCCAACCTGAGCCCCAGAGAGGA ACCCGACGACATCGACTGCTGGTGCTACGGCGTGGAAAACGTGCGG GTGGCCTACGGCAAGTGCGATAGCGCCGGCAGAAGCAGAAGAAGCA GGCGGGCCATCGACCTGCCCACCCACGAAAACCACGGCCTGAAAAC CCGGCAGGAAAAGTGGATGACCGGCCGGATGGGCGAGCGGCAGCTG CAGAAAATTGAGCGGTGGTTTGTGCGGAACCCCTTCTTCGCCGTGA CCGCCCTGACAATCGCCTACCTCGTGGGCAGCAACATGACCCAGAG AGTCGTGATCGCCCTGCTGGTGCTGGCTGTGGGCCCTGCCTATAGC GCCATTGATCTGCCTACACACGAGAATCATGGGCTGAAAACAAGAC AGGAAAAATGGATGACTGGGCGCATGGGAGAAAGACAGCTGCAGAA AATCGAACGGTGGTTCGTGCGCAATCCTTTTTTTGCTGTGACTGCT CTGACCATTGCCTATCTCGTGGGATCCAATATGACACAGCGGGTCG TGATTGCTCTGCTGGTGCTGGCAGTGGGACCCGCTTACTCCGCCCA CTGTATCGGCATCACCGACCGGGACTTCATCGAGGGCGTGCACGGC GGAACATGGGTGTCCGCTACCCTGGAACAGGATAAGTGCGTGACCG TGATGGCCCCCGACAAGCCCAGCCTGGACATCAGCCTGGAAACCGT GGCCATCGATAGACCCGCCGAAGTGCGGAAAGTGTGCTACAACGCC GTGCTGACCCACGTGAAGATCAACGACAAGTGCCCCAGCACCGGCG AAGCCCACCTGGCCGAAGAGAACGAGGGCGACAACGCCTGCAAGCG GACCTACAGCGATAGAGGCTGGGGCAATGGCTGCGGCCTGTTTGGC AAGGGCAGCATCGTGGCCTGCGCCAAGTTCACCTGTGCCAAGAGCA TGAGCCTGTTCGAGGTGGACCAGACCAAGATCCAGTACGTGATCCG GGCTCAGCTGCACGTGGGCGCCAAGCAGGAAAACTGGAACACCGAC ATCAAGACCCTGAAGTTCGACGCCCTGAGCGGCTCCCAGGAAGTGG AATTCATCGGCTATGGCAAGGCCACCCTGGAATGTCAGGTGCAGAC CGCCGTGGACTTCGGCAACAGCTATATCGCCGAGATGGAAACCGAG AGCTGGATCGTGGACCGGCAGTGGGCCCAGGATCTGACACTGCCTT GGCAGTCTGGCTCTGGCGGAGTGTGGCGGGAAATGCACCACCTGGT GGAATTCGAGCCTCCCCACGCCGCCACCATTAGAGTGCTGGCCCTG GGCAATCAGGAAGGCTCTCTGAAAACAGCCCTGACCGGCGCCATGA GAGTGACCAAGGACACCAACGACAACAACCTGTACAAGCTGCACGG GGGGCACGTGTCCTGCAGAGTGAAACTGTCTGCCCTGACACTGAAG GGCACCAGCTACAAGATCTGCACCGACAAGATGTTCTTCGTGAAGA ACCCCACCGACACCGGCCACGGCACAGTCGTGATGCAAGTGAAGGT GTCCAAGGGCGCTCCCTGCCGGATCCCTGTGATCGTGGCCGATGAT CTGACAGCCGCCATCAACAAGGGCATCCTCGTGACAGTAACCCTAT CGCCTCCACCAACGATGACGAGGTGCTGATCGAAGTGAACCCCCCC TTCGGCGACTCCTACATCATCGTGGGACGGGGCGACAGCAGACTGA CCTACCAGTGGCACAAAGAGGGCAGCAGCATCGGCAAGCTGTTCAC CCAGACCATGAAGGGCGTGGAACGGCTGGCCGTGATGGGAGATACC GCCTGGGATTTCTCTAGCGCTGGCGGCTTCTTCACCAGCGTGGGCA AGGGAATCCACACCGTGTTCGGCAGCGCCTTCCAGGGACTGTTCGG CGGCCTGAACTGGATCACCAAAGTGATCATGGGCGCTGTGCTGATC TGGGTGGGAATCAACACCCGGAACATGACCATGAGCATGTCCATGA TCCTCGTGGGAGTGATTATGATGTTCCTGAGCCTGGGCGTGGGTGC C YF_Sen2000_ ATGTGGCTGGTGTCCCTGGCCATCGTGACAGCCTGTGCTGGCGCTG 70 JEVsp_N153T TGACACTCGTGCGGAAGAACAGATGGCTGCTGCTGAACGTGACCAG CGAGGACCTGGGCAAGACCTTCTCTGTGGGCACCGGCAACTGCACC ACCAACATCCTGGAAGCCAAGTACTGGTGCCCCGACAGCATGGAGT ACAACTGCCCCAACCTGAGCCCCAGAGAGGAACCCGACGACATCGA CTGCTGGTGCTACGGCGTGGAAAACGTGCGGGTGGCCTACGGCAAG TGCGATAGCGCCGGCAGAAGCAGAAGAAGCAGGCGGGCCATCGACC TGCCCACCCACGAAAACCACGGCCTGAAAACCCGGCAGGAAAAGTG GATGACCGGCCGGATGGGCGAGCGGCAGCTGCAGAAAATCGAGAGA TGGCTCGTGCGCAACCCCTTCTTCGCCGTGACCGCCCTGACAATCG CCTACCTCGTGGGCAGCAACATGACCCAGAGAGTCGTGATCGCCCT GCTGGTGCTGGCTGTGGGCCCTGCCTATAGCGCCCACTGTATCGGC ATCACCGACCGGGACTTCATCGAGGGCGTGCACGGCGGAACATGGG TGTCCGCTACCCTGGAACAGGATAAGTGCGTGACCGTGATGGCCCC CGACAAGCCCAGCCTGGACATCAGCCTGGAAACCGTGGCCATTGAC GGACCCGCCGAGGCCAGAAAAGTGTGCTACAACGCCGTGCTGACCC ACGTGAAGATCAACGACAAGTGCCCCAGCACCGGCGAAGCCCACCT GGCCGAAGAGAACGAGGGCGACAACGCCTGCAAGCGGACCTACAGC GATAGAGGCTGGGGCAATGGCTGCGGCCTGTTTGGCAAGGGCAGCA TCGTGGCCTGCGCCAAGTTCACCTGTGCCAAGAGCATGAGCCTGTT CGAGGTGGACCAGACCAAGATCCAGTACGTGATCCGGGCCCAGCTG CACGTGGGAGCCAAGCAGGAAAACTGGACCACCGACATCAAGACCC TGAAGTTCGACGCCCTGAGCGGCTCCCAGGAAGCCGAGTTTACCGG CTATGGCAAGGCCACCCTGGAATGCCAGGTGCAGACCGCCGTGGAC TTCGGCAACAGCTATATCGCCGAGATGGAAAAAGAAAGCTGGATCG TGGACCGGCAGTGGGCCCAGGACCTGACACTGCCTTGGCAGTCTGA TCTGGCGGCGTGTGGCGGGAAATGCACCACCTGGTGGAATTCGAGC CTCCCCATGCCGCCACCATCAGAGTGCTGGCCCTGGGCAATCAGGA AGGCTCTCTGAAAACAGCCCTGACCGGCGCCATGAGAGTGACCAAG GACACCAACGACAACAACCTGTACAAGCTGCATGGCGGCCACGTGT CCTGCAGAGTGAAGCTGTCTGCCCTGACCCTGAAAGGCACCAGCTA CAAGATGTGCACCGACAAGATGAGCTTCGTGAAGAACCCCACCGAC ACCGGCCACGGCACAGTCGTGATGCAAGTGAAGGTGCCCAAGGGCG CTCCCTGCAGAATCCCTGTGATCGTGGCCGATGATCTGACAGCCGC CATCAACAAGGGCATCCTCGTGACAGTGAACCCTATCGCCTCCACC AACGATGACGAGGTGCTGATCGAAGTGAACCCCCCCTTCGGCGACT CCTACATCATCGTGGGCACAGGCGACAGCAGACTGACCTACCAGTG GCACAAAGAGGGCAGCAGCATCGGCAAGCTGTTCACCCAGACCATG AAGGGCGCCGAGAGACTGGCTGTGATGGGAGATGCCGCCTGGGACT TTAGCAGCGCTGGCGGCTTCTTTACCAGCGTGGGCAAGGGAATCCA CACCGTGTTCGGCAGCGCCTTCCAGGGACTGTTTGGCGGCCTGAGC TGGATCACCAAAGTGATCATGGGCGCTGTGCTGATCTGGGTGGGAA TCAACACCCGGAACATGACCATGAGCATGTCCATGATCCTCGTGGG AGTGATTATGATGTTCCTGAGCCTGGGCGTGGGCGCC YF_17D_IgGksp_SE ATGGAAACCCCTGCCCAGCTGCTGTTCCTGCTGCTGCTGTGGCTGC 71 CTGATACCACCGGCGTGACACTCGTGCGGAAGAACAGATGGCTGCT GCTGAACGTGACCAGCGAGGACCTGGGCAAGACCTTCTCTGTGGGC ACCGGCAACTGCACCACCAACATCCTGGAAGCCAAGTACTGGTGCC CCGACAGCATGGAGTACAACTGCCCCAACCTGAGCCCCAGAGAGGA ACCCGACGACATCGACTGCTGGTGCTACGGCGTGGAAAACGTGCGG GTGGCCTACGGCAAGTGCGATAGCGCCGGCAGAAGCAGAAGAAGCA GGCGGGCCATCGACCTGCCCACCCACGAAAACCACGGCCTGAAAAC CCGGCAGGAAAAGTGGATGACCGGCCGGATGGGCGAGCGGCAGCTG CAGAAAATTGAGCGGTGGTTTGTGCGGACCCCTTCTTCGCCGTGAC CGCCCTGACAATCGCCTACCTCGTGGGCAGCAACATGACCCAGAGA GTCGTGATCGCCCTGCTGGTGCTGGCTGTGGGCCCTGCCTATAGCG CCATTGATCTGCCTACACACGAGAATCATGGGCTGAAAACAAGACA GGAAAAATGGATGACTGGGCGCATGGGAGAAAGACAGCTGCAGAAA ATCGAACGGTGGTTCGTGCGCAATCCTTTTTTTGCTGTGACTGCTC TGACCATTGCCTATCTCGTGGGATCCAATATGACACAGCGGGTCGT GATTGCTCTGCTGGTGCTGGCAGTGGGACCCGCTTACTCCGCCCAC TGTATCGGCATCACCGACCGGGACTTCATCGAGGGCGTGCACGGCG GAACATGGGTGTCCGCTACCCTGGAACAGGATAAGTGCGTGACCGT GATGGCCCCCGACAAGCCCAGCCTGGACATCAGCCTGGAAACCGTG GCCATCGATAGACCCGCCGAAGTGCGGAAAGTGTGCTACAACGCCG TGCTGACCCACGTGAAGATCAACGACAAGTGCCCCAGCACCGGCGA AGCCCACCTGGCCGAAGAGAACGAGGGCGACAACGCCTGCAAGCGG ACCTACAGCGATAGAGGCTGGGGCAATGGCTGCGGCCTGTTTGGCA AGGGCAGCATCGTGGCCTGCGCCAAGTTCACCTGTGCCAAGAGCAT GAGCCTGTTCGAGGTGGACCAGACCAAGATCCAGTACGTGATCCGG GCTCAGCTGCACGTGGGCGCCAAGCAGGAAAACTGGAACACCGACA TCAAGACCCTGAAGTTCGACGCCCTGAGCGGCTCCCAGGAAGTGGA ATTCATCGGCTATGGCAAGGCCACCCTGGAATGTCAGGTGCAGACC GCCGTGGACTTCGGCAACAGCTATATCGCCGAGATGGAAACCGAGA GCTGGATCGTGGACCGGCAGTGGGCCCAGGATCTGACACTGCCTTG GCAGTCTGGCTCTGGCGGAGTGTGGCGGGAAATGCACCACCTGGTG GAATTCGAGCCTCCCCACGCCGCCACCATTAGAGTGCTGGCCCTGG GCAATCAGGAAGGCTCTCTGAAAACAGCCCTGACCGGCGCCATGAG AGTGACCAAGGACACCAACGACAACAACCTGTACAAGCTGCACGGG GGGCACGTGTCCTGCAGAGTGAAACTGTCTGCCCTGACACTGAAGG GCACCAGCTACAAGATCTGCACCGACAAGATGTTCTTCGTGAAGAA CCCCACCGACACCGGCCACGGCACAGTCGTGATGCAAGTGAAGGTG TCCAAGGGCGCTCCCTGCCGGATCCCTGTGATCGTGGCCGATGATC TGACAGCCGCCATCAACAAGGGCATCCTCGTGACAGTGAACCCTAT CGCCTCCACCAACGATGACGAGGTGCTGATCGAAGTGAACCCCCCC TTCGGCGACTCCTACATCATCGTGGGACGGGGCGACAGCAGACTGA CCTACCAGTGGCACAAAGAGGGCAGCAGCATCGGCAAGCTGTTCAC CCAGACCATGAAGGGCGTGGAACGGCTGGCCGTGATGGGAGATACC GCCTGGGATTTCTCTAGCGCTGGCGGCTTCTTCACCAGCGTGGGCA AGGGAATCCACACCGTGTTCGGCAGCGCCTTCCAGGGACTGTTCGG CGGCCTGAACTGGATCACCAAAGTGATCATGGGCGCTGTGCTGATC TGGGTGGGAATCAACACCCGGAACATGACCATGAGCATGTCCATGA TCCTCGTGGGAGTGATTATGATGTTCCTGAGCCTGGGCGTGGGTGC C YF_17D_JEVsp_SE ATGTGGCTGGTGTCCCTGGCCATCGTGACAGCCTGTGCTGGCGCTG 72 TGACACTCGTGCGGAAAAACAGATGGCTGCTGCTGAACGTGACCAG CGAGGACCTGGGCAAGACCTTCTCTGTAGGCACCGGCAACTGCACC ACCAACATCCTGGAAGCCAAGTACTGGTGCCCCGACAGCATGGAGT ACAACTGCCCCAACCTGAGCCCCAGAGAGGAACCCGACGACATCGA CTGCTGGTGCTACGGCGTGGAAAACGTGCGGGTGGCCTACGGCAAG TGCGATAGCGCCGGCAGAAGCAGAAGAAGCAGGCGGGCCATCGACC TGCCCACCCACGAAAACCACGGCCTGAAAACCCGGCAGGAAAAGTG GATGACCGGCCGGATGGGCGAGCGGCAGCTGCAGAAAATTGAGCGG TGGTTTGTGCCGGAACCCCTTCTTCGCCGTGACCGCCCTGACAATC GCCTACCTCGTGGGCAGCAACATGACCCAGAGAGTCGTGATCGCCC TGCTGGTGCTGGCTGTGGGCCCTGCCTATAGCGCCATTGATCTGCC TACACACGAGAATCATGGGCTGAAAACAAGACAGGAAAAATGGATG ACTGGGCGCATGGGAGAAAGACAGCTGCAGAAAATCGAACGGTGGT TCGTGCGCAATCCTTTTTTTGCTGTGACTGCTCTGACCATTGCCTA TCTCGTGGGATCCAATATGACACAGCGGGTCGTGATTGCTCTGCTG GTGCTGGCAGTGGGACCCGCTTACTCCGCCCACTGTATCGGCATCA CCGACCGGGACTTCATCGAGGGCGTGCACGGCGGAACATGGGTGTC CGCTACCCTGGAACAGGATAAGTGCGTGACCGTGATGGCCCCCGAC AAGCCCAGCCTGGACATCAGCCTGGAAACCGTGGCCATCGATAGAC CCGCCGAAGTGCGGAAAGTGTGCTACAACGCCGTGCTGACCCACGT GAAGATCAACGACAAGTGCCCCAGCACCGGCGAAGCCCACCTGGCC GAAGAGAACGAGGGCGACAACGCCTGCAAGCGGACCTACAGCGATA GAGGCTGGGGCAATGGCTGCGGCCTGTTTGGCAAGGGCAGCATCGT GGCCTGCGCCAAGTTCACCTGTGCCAAGAGCATGAGCCTGTTCGAG GTGGACCAGACCAAGATCCAGTACGTGATCCGGGCCCAGCTGCACG TGGGAGCCAAGCAGGAAAACTGGAACACCGACATCAAGACCCTGAA GTTCGACGCCCTGAGCGGCTCCCAGGAAGTGGAATTCATCGGCTAT GGCAAGGCCACCCTGGAATGTCAGGTGCAGACCGCCGTGGACTTCG GCAACAGCTATATCGCCGAGATGGAAACCGAGAGCTGGATCGTGGA CCGGCAGTGGGCTCAGGATCTGACCCTGCCTTGGCAGTCTGGCTCT GGCGGAGTGTGGCGGGAAATGCACCACCTGGTGGAATTCGAGCCTC CCCACGCCGCCACCATTAGAGTGCTGGCCCTGGGCAATCAGGAAGG CTCTCTGAAAACAGCCCTGACCGGCGCCATGAGAGTGACCAAGGAC ACCAACGACAACAACCTGTACAAGCTGCATGGCGGCCACGTGTCCT GCAGAGTGAAGCTGTCTGCCCTGACACTGAAGGGCACCAGCTACAA GATCTGCACCGACAAGATGTTCTTCGTGAAGAACCCCACCGACACC GGCCACGGCACAGTCGTGATGCAAGTGAAGGTGTCCAAGGGCGCTC CCTGCCGGATCCCTGTGATCGTGGCCGATGATCTGACAGCCGCCAT CAACAAGGGCATCCTCGTGACAGTGAACCCTATCGCCTCCACCAAC GATGACGAGGTGCTGATCGAAGTGAACCCCCCCTTCGGCGACTCCT ACATCATCGTGGGACGGGGCGACAGCAGACTGACCTACCAGTGGCA CAAAGAGGGCAGCAGCATCGGCAAGCTGTTCACCCAGACCATGAAG GGCGTGGAACGGCTGGCCGTGATGGGAGATACCGCCTGGGATTTCT CTAGCGCTGGCGGCTTCTTCACCAGCGTGGGCAAGGGAATCCACAC CGTGTTCGGCAGCGCCTTCCAGGGACTGTTCGGCGGCCTGAACTGG ATCACCAAAGTGATCATGGGCGCTGTGCTGATCTGGGTGGGAATCA ACACCCGGAACATGACCATGAGCATGTCCATGATCCTCGTGGGAGT GATTATGATGTTCCTGAGCCTGGGTGTGGGCGCC YF_SEN2000_ ATGTGGCTGGTGTCCCTGGCCATCGTGACAGCCTGTGCTGGCGCTG 73 prME_JEVsp TGACACTCGTGCGGAAGAACAGATGGCTGCTGCTGAACGTGACCAG CGAGGACCTGGGCAAGACCTTCTCTGTGGGCACCGGCAACTGCACC ACCAACATCCTGGAAGCCAAGTACTGGTGCCCCGACAGCATGGAGT ACAACTGCCCCAACCTGAGCCCCAGAGAGGAACCCGACGACATCGA CTGCTGGTGCTACGGCGTGGAAAACGTGCGGGTGGCCTACGGCAAG TGCGATAGCGCCGGCAGAAGCAGAAGAAGCAGGCGGGCCATCGACC TGCCCACCCACGAAAACCACGGCCTGAAAACCCGGCAGGAAAAGTG GATGACCGGCCGGATGGGCGAGCGGCAGCTGCAGAAAATCGAGAGA TGGCTCGTGCGCAACCCCTTCTTCGCCGTGACCGCCCTGACAATCG CCTACCTCGTGGGCAGCAACATGACCCAGAGAGTCGTGATCGCCCT GCTGGTGCTGGCTGTGGGCCCTGCCTATAGCGCCCACTGTATCGGC ATCACCGACCGGGACTTCATCGAGGGCGTGCACGGCGGAACATGGG TGTCCGCTACCCTGGAACAGGATAAGTGCGTGACCGTGATGGCCCC CGACAAGCCCAGCCTGGACATCAGCCTGGAAACCGTGGCCATTGAC GGACCCGCCGAGGCCAGAAAAGTGTGCTACAACGCCGTGCTGACCC ACGTGAAGATCAACGACAAGTGCCCCAGCACCGGCGAAGCCCACCT GGCCGAAGAGAACGAGGGCGACAACGCCTGCAAGCGGACCTACAGC GATAGAGGCTGGGGCAATGGCTGCGGCCTGTTTGGCAAGGGCAGCA TCGTGGCCTGCGCCAAGTTCACCTGTGCCAAGAGCATGAGCCTGTT CGAGGTGGACCAGACCAAGATCCAGTACGTGATCCGGGCCCAGCTG CACGTGGGAGCCAAGCAGGAAAACTGGAACACCGACATCAAGACCC TGAAGTTCGACGCCCTGAGCGGCTCCCAGGAAGCCGAGTTTACCGG CTATGGCAAGGCCACCCTGGAATGCCAGGTGCAGAACGCCGTGGAC TTCGGCAACAGCTATATCGCCGAGATGGAAAAAGAAAGCTGGATCG TGGACCGGCAGTGGGCCCAGGACCTGACACTGCCTTGGCAGTCTGG ATCTGGCGGCGTGTGGCGGGAAATGCACCACCTGGTGGAATTCGAG CCTCCCCATGCCGCCACCATCAGAGTGCTGGCCCTGGGCAATCAGG AAGGCTCTCTGAAAACAGCCCTGACCGGCGCCATGAGAGTGACCAA GGACACCAACGACAACAACCTGTACAAGCTGCATGGCGGCCACGTG TCCTGCAGAGTGAAGCTGTCTGCCCTGACCCTGAAAGGCACCAGCT ACAAGATGTGCACCGACAAGATGAGCTTCGTGAAGAACCCCACCGA CACCGGCCACGGCACAGTCGTGATGCAAGTGAAGGTGCCCAAGGGC GCTCCCTGCAGAATCCCTGTGATCGTGGCCGATGATCTGACAGCCG CCATCAACAAGGGCATCCTCGTGACAGTGAACCCTATCGCCTCCAC CAACGATGACGAGGTGCTGATCGAAGTGAACCCCCCCTTCGGCGAC TCCTACATCATCGTGGGCACAGGCGACAGCAGACTGACCTACCAGT GGCACAAAGAGGGCAGCAGCATCGGCAAGCTGTTCACCCAGACCAT GAAGGGCGCCGAGAGACTGGCTGTGATGGGAGATGCCGCCTGGGAC TTTAGCAGCGCTGGCGGCTTCTTTACCAGCGTGGGCAAGGGAATCC ACACCGTGTTCGGCAGCGCCTTCCAGGGACTGTTTGGCGGCCTGAG CTGGATCACCAAAGTGATCATGGGCGCTGTGCTGATCTGGGTGGGA ATCAACACCCGGCAACATGACCATGAGCATGTCCATGATCCTCGTG GGAGTGATTATGATGTTCCTGAGCCTGGGCGTGGGCGCC 74 YF_Sen2000_ ATGGAAACCCCTGCCCAGCTGCTGTTCCTGCTGCTGCTGTGGCTGC prME_IgGksp CTGATACCACCGGCGTGACACTCGTGCGGAAGAACAGATGGCTGCT GCTGAACGTGACCAGCGAGGACCTGGGCAAGACCTTCTCTGTGGGC ACCGGCAACTGCACCACCAACATCCTGGAAGCCAAGTACTGGTGCC CCGACAGCATGGAGTACAACTGCCCCAACCTGAGCCCCAGAGAGGA ACCCGACGACATCGACTGCTGGTGCTACGGCGTGGAAAACGTGCGG GTGGCCTACGGCAAGTGCGATAGCGCCGGCAGAAGCAGAAGAAGCA GGCGGGCCATCGACCTGCCCACCCACGAAAACCACGGCCTGAAAAC CCGGCAGGAAAAGTGGATGACCGGCCGGATGGGCGAGCGGCAGCTG CAGAAAATCGAGAGATGGCTCGTGCGCAACCCCTTCTTCGCCGTGA CCGCCCTGACAATCGCCTACCTCGTGGGCAGCAACATGACCCAGAG AGTCGTGATCGCCCTGCTGGTGCTGGCTGTGGGCCCTGCCTATAGC GCCCACTGTATCGGCATCACCGACCGGGACTTCATCGAGGGCGTGC ACGGCGGAACATGGGTGTCCGCTACCCTGGAACAGGATAAGTGCGT GACCGTGATGGCCCCCGACAAGCCCAGCCTGGACATCAGCCTGGAA ACCGTGGCCATTGACGGACCCGCCGAGGCCAGAAAAGTGTGCTACA ACGCCGTGCTGACCCACGTGAAGATCAACGACAAGTGCCCCAGCAC CGGCGAAGCCCACCTGGCCGAAGAGAACGAGGGCGACAACGCCTGC AAGCGGACCTACAGCGATAGAGGCTGGGGCAATGGCTGCGGCCTGT TTGGCAAGGGCAGCATCGTGGCCTGCGCCAAGTTCACCTGTGCCAA GAGCATGAGCCTGTTCGAGGTGGACCAGACCAAGATCCAGTACGTG ATCCGGGCTCAGCTGCACGTGGGCGCCAAGCAGGAAAACTGGAACA CCGACATCAAGACCCTGAAGTTCGACGCCCTGAGCGGCTCCCAGGA AGCCGAGTTTACCGGCTATGGCAAGGCCACCCTGGAATGCCAGGTG CAGACCGCCGTGGACTTCGGCAACAGCTATATCGCCGAGATGGAAA AAGAAAGCTGGATCGTGGACCGGCAGTGGGCCCAGGATCTGACACT GCCTTGGCAGTCTGGCTCTGGCGGAGTGTGGCGGGAAATGCACCAC CTGGTGGAATTCGAGCCTCCCCACGCCGCCACCATTAGAGTGCTGG CCCTGGGCAATCAGGAAGGCTCTCTGAAAACAGCCCTGACCGGCGC CATGAGAGTGACCAAGGACACCAACGACAACAACCTGTACAAGCTG CACGGGGGGCACGTGTCCTGCAGAGTGAAACTGTCTGCCCTGACAC TGAAGGGCACCAGCTACAAGATGTGCACCGACAAGATGAGCTTCGT GAAGAACCCCACCGACACCGGCCACGGCACAGTCGTGATGCAAGTG AAGGTGCCCAAGGGCGCTCCCTGCAGAATCCCTGTGATCGTGGCCG ATGATCTGACAGCCGCCATCAACAAGGGCATCCTCGTGACAGTGAA CCCTATCGCCTCCACCAACGATGACGAGGTGCTGATCGAAGTGAAC CCCCCCTTCGGCGACTCCTACATCATCGTGGGCACAGGCGACAGCA GACTGACCTACCAGTGGCACAAAGAGGGCAGCAGCATCGGCAAGCT GTTCACCCAGACCATGAAGGGCGCCGAGAGACTGGCTGTGATGGGA GATGCCGCCTGGGACTTTAGCAGCGCTGGCGGCTTCTTTACCAGCG TGGGCAAGGGAATCCACACCGTGTTCGGCAGCGCCTTCCAGGGACT GTTTGGCGGCCTGAGCTGGATCACCAAAGTGATCATGGGCGCTGTG CTGATCTGGGTGGGAATCAACACCCGGAACATGACCATGAGCATGT CCATGATCCTCGTGGGGAGTGATTATGATGTTCCTGAGCCTGGGCG TGGGAGCC YF_17D_JEVsp_ ATGTGGCTGGTGTCCCTGGCCATCGTGACAGCCTGTGCTGGCGCTG 75 SE_mutFL TGACACTCGTGCGGAAAAACAGATGGCTGCTGCTGAACGTGACCAG CGAGGACCTGGGCAAGACCTTCTCTGTAGGCACCGGCAACTGCACC ACCAACATCCTGGAAGCCAAGTACTGGTGCCCCGACAGCATGGAGT ACAACTGCCCCAACCTGAGCCCCAGAGAGGAACCCGACGACATCGA CTGCTGGTGCTACGGCGTGGAAAACGTGCGGGTGGCCTACGGCAAG TGCGATAGCGCCGGCAGAAGCAGAAGAAGCAGGCGGGCCATCGACC TGCCCACCCACGAAAACCACGGCCTGAAAACCCGGCAGGAAAAGTG GATGACCGGCCGGATGGGCGAGCGGCAGCTGCAGAAAATTGAGCGG TGGTTTGTGCGGAACCCCTTCTTCGCCGTGACCGCCCTGACAATCG CCTACCTCGTGGGCAGCAACATGACCCAGAGAGTCGTGATCGCCCT GCTGGTGCTGGCTGTGGGCCCTGCCTATAGCGCCATTGATCTGCCT ACACACGAGAATCATGGGCTGAAAACAAGACAGGAAAAATGGATGA CTGGGCGCATGGGAGAAAGACAGCTGCAGAAAATCGAACGGTGGTT CGTGCGCAATCCTTTTTTTGCTGTGACTGCTCTGACCATTGCCTAT CTCGTGGGATCCAATATGACACAGCGGGTCGTGATTGCTCTGCTGG TGCTGGCAGTGGGACCCGCTTACTCCGCCCACTGTATCGGCATCAC CGACCGGGACTTCATCGAGGGCGTGCACGGCGGAACATGGGTGTCC GCTACCCTGGAACAGGATAAGTGCGTGACCGTGATGGCCCCCGACA AGCCCAGCCTGGACATCAGCCTGGAAACCGTGGCCATCGATAGACC CGCCGAAGTGCGGAAAGTGTGCTACAACGCCGTGCTGACCCACGTG AAGATCAACGACAAGTGCCCCAGAGAAGGCGAAGCCCACCTGGCCG AAGAGAACGAGGGCGACAACGCCTGCAAGCGGACCTACAGCGATAG AGGCAGAGGCAATGGCTGCGGCAGATTTGGCAAGGGCAGCATCGTG GCCTGCGCCAAGTTCACCTGTGCCAAGAGCATGAGCCTGTTCGAGG TGGACCAGACCAAGATCCAGTACGTGATCCGGGCCCAGCTGCACGT GGGAGCCAAGCAGGAAAACTGGAACACCGACATCAAGACCCTGAAG TTCGACGCCCTGAGCGGCTCCCAGGAAGTGGAATTCATCGGCTATG GCAAGGCCACCCTGGAATGTCAGGTGCAGACCGCCGTGGACTTCGG CAACAGCTATATCGCCGAGATGGAAACCGAGAGCTGGATCGTGGAC CGGCAGTGGGCTCAGGATCTGACCCTGCCTTGGCAGTCTGGCTCTG GCGGAGTGTGGCGGGAAATGCACCACCTGGTGGAATTCGAGCCTCC CCACGCCGCCACCATTAGAGTGCTGGCCCTGGGCAATCAGGAAGGC TCTCTGAAAACAGCCCTGACCGGCGCCATGAGAGTGACCAAGGACA CCAACGACAACAACCTGTACAAGCTGCATGGCGGCCACGTGTCCTG CAGAGTGAAGCTGTCTGCCCTGACACTGAAGGGCACCAGCTACAAG ATCTGCACCGACAAGATGTTCTTCGTGAAGAACCCCACCGACACCG GCCACGGCACAGTCGTGATGCAAGTGAAGGTGTCCAAGGGCGCTCC CTGCCGGATCCCTGTGATCGTGGCCGATGATCTGACAGCCGCCATC AACAAGGGCATCCTCGTGACAGTGAACCCTATCGCCTCCACCAACG ATGACGAGGTGCTGATCGAAGTGAACCCCCCCTTCGGCGACTCCTA CATCATCGTGGGACGGGGCGACAGCAGACTGACCTACCAGTGGCAC AAAGAGGGCAGCAGCATCGGCAAGCTGTTCACCCAGACCATGAAGG GCGTGGAACGGCTGGCCGTGATGGGAGATACCGCCTGGGATTTCTC TAGCGCTGGCGGCTTCTTCACCAGCGTGGGCAAGGGAATCCACACC GTGTTCGGCAGCGCCTTCCAGGGACTGTTCGGCGGCCTGAACTGGA TCACCAAAGTGATCATGGGCGCTGTGCTGATCTGGGTGGGAATCAA CACCCGGAACATGACCATGAGCATGTCCATGATCCTCGTGGGAGTG ATTATGATGTTCCTGAGCCTGGGTGTGGGCGCC YF_17D_IgGksp_ ATGGAAACCCCTGCCCAGCTGCTGTTCCTGCTGCTGCTGTGGCTGC 76 SE_mutFL CTGATACCACCGGCGTGACACTCGTGCGGAAGAACAGATGGCTGCT GCTGAACGTGACCAGCGAGGACCTGGGCAAGACCTTCTCTGTGGGC ACCGGCAACTGCACCACCAACATCCTGGAAGCCAAGTACTGGTGCC CCGACAGCATGGAGTACAACTGCCCCAACCTGAGCCCCAGAGAGGA ACCCGACGACATCGACTGCTGGTGCTACGGCGTGGAAAACGTGCGG GTGGCCTACGGCAAGTGCGATAGCGCCGGCAGAAGCAGAAGAAGCA GGCGGGCCATCGACCTGCCCACCCACGAAAACCACGGCCTGAAAAC CCGGCAGGAAAAGTGGATGACCGGCCGGATGGGCGAGCGGCAGCTG CAGAAAATTGAGCGGTGGTTTGTGCGGAACCCCTTCTTCGCCGTGA CCGCCCTGACAATCGCCTACCTCGTGGGCAGCAACATGACCCAGAG AGTCGTGATCGCCCTGCTGGTGCTGGCTGTGGGCCCTGCCTATAGC GCCATTGATCTGCCTACACACGAGAATCATGGGCTGAAAACAAGAC AGGAAAAATGGATGACTGGGCGCATGGGAGAAAGACAGCTGCAGAA AATCGAACGGTGGTTCGTGCGCAATCCTTTTTTTGCTGTGACTGCT CTGACCATTGCCTATCTCGTGGGATCCAATATGACACAGCGGGTCG TGATTGCTCTGCTGGTGCTGGCAGTGGGACCCGCTTACTCCGCCCA CTGTATCGGCATCACCGACCGGGACTTCATCGAGGGCGTGCACGGC GGAACATGGGTGTCCGCTACCCTGGAACAGGATAAGTGCGTGACCG TGATGGCCCCCGACAAGCCCAGCCTGGACATCAGCCTGGAAACCGT GGCCATCGATAGACCCGCCGAAGTGCGGAAAGTGTGCTACAACGCC GTGCTGACCCACGTGAAGATCAACGACAAGTGCCCCAGAGAAGGCG AAGCCCACCTGGCCGAAGAGAACGAGGGCGACAACGCCTGCAAGCG GACCTACAGCGATAGAGGCAGAGGCAATGGCTGCGGCAGATTTGGC AAGGGCAGCATCGTGGCCTGCGCCAAGTTCACCTGTGCCAAGAGCA TGAGCCTGTTCGAGGTGGACCAGACCAAGATCCAGTACGTGATCCG GGCTCAGCTGCACGTGGGCGCCAAGCAGGAAAACTGGAACACCGAC ATCAAGACCCTGAAGTTCGACGCCCTGAGCGGCTCCCAGGAAGTGG AATTCATCGGCTATGGCAAGGCCACCCTGGAATGTCAGGTGCAGAC CGCCGTGGACTTCGGCAACAGCTATATCGCCGAGATGGAAACCGAG AGCTGGATCGTGGACCGGCAGTGGGCCCAGGATCTGACACTGCCTT GGCAGTCTGGCTCTGGCGGAGTGTGGCGGGAAATGCACCACCTGGT GGAATTCGAGCCTCCCCACGCCGCCACCATTAGAGTGCTGGCCCTG GGCAATCAGGAAGGCTCTCTGAAAACAGCCCTGACCGGCGCCATGA GAGTGACCAAGGACACCAACGACAACAACCTGTACAAGCTGCACGG GGGGCACGTGTCCTGCAGAGTGAAACTGTCTGCCCTGACACTGAAG GGCACCAGCTACAAGATCTGCACCGACAAGATGTTCTTCGTGAAGA ACCCCACCGACACCGGCCACGGCACAGTCGTGATGCAAGTGAAGGT GTCCAAGGGCGCTCCCTGCCGGATCCCTGTGATCGTGGCCGATGAT CTGACAGCCGCCATCAACAAGGGCATCCTCGTGACAGTGAACCCTA TCGCCTCCACCAACGATGACGAGGTGCTGATCGAAGTGAACCCCCC CTTCGGCGACTCCTACATCATCGTGGGACGGGGCGACAGCAGACTG ACCTACCAGTGGCACAAAGAGGGCAGCAGCATCGGCAAGCTGTTCA CCCAGACCATGAAGGGCGTGGAACGGCTGGCCGTGATGGGAGATAC CGCCTGGGATTTCTCTAGCGCTGGCGGCTTCTTCACCAGCGTGGGC AAGGGAATCCACACCGTGTTCGGCAGCGCCTTCCAGGGACTGTTCG GCGGCCTGAACTGGATCACCAAAGTGATCATGGGCGCTGTGCTGAT CTGGGTGGGAATCAACACCCGGAACATGACCATGAGCATGTCCATG ATCCTCGTGGGAGTGATTATGATGTTCCTGAGCCTGGGCGTGGGTG CC YF_17D_JEVsp_ ATGTGGCTGGTGTCCCTGGCCATCGTGACAGCCTGTGCTGGCGCTG 77 SE_no_polyNs_ TGACACTCGTGCGGAAGAACAGATGGCTGCTGCTGAACGTGACCAG mutFL CGAGGACCTGGGCAAGACCTTCTCTGTAGGCACCGGCAACTGCACC ACCAACATCCTGGAAGCCAAGTACTGGTGCCCCGACAGCATGGAGT ACAACTGCCCCAACCTGAGCCCCAGAGAGGAACCCGACGACATCGA CTGCTGGTGCTACGGCGTGGAAAACGTGCGGGTGGCCTACGGCAAG TGCGATAGCGCCGGCAGAAGCAGAAGAAGCAGGCGGGCCATCGACC TGCCCACCCACGAAAACCACGGCCTGAAAACCCGGCAGGAAAAGTG GATGACCGGCCGGATGGGCGAGCGGCAGCTGCAGAAAATTGAGCGG TGGTTTGTGCGGAACCCCTTCTTCGCCGTGACCGCCCTGACAATCG CCTACCTCGTGGGCAGCAACATGACCCAGAGAGTCGTGATCGCCCT GCTGGTGCTGGCTGTGGGCCCTGCCTATAGCGCCATTGATCTGCCT ACACACGAGAATCATGGGCTGAAAACAAGACAGGAGAAATGGATGA CTGGGCGCATGGGAGAAAGACAGCTGCAGAAAATCGAACGGTGGTT CGTGCGCAATCCTTTCTTTGCTGTGACTGCTCTGACCATTGCCTAT CTCGTGGGATCCAATATGACACAGCGGGTCGTGATTGCTCTGCTGG TGCTGGCAGTGGGACCCGCTTACTCCGCCCACTGTATCGGCATCAC CGACCGGGACTTCATCGAGGGCGTGCACGGCGGAACATGGGTGTCC GCTACCCTGGAACAGGATAAGTGCGTGACCGTGATGGCCCCCGACA AGCCCAGCCTGGACATCAGCCTGGAAACCGTGGCCATCGATAGACC CGCCGAAGTGCGGAAAGTGTGCTACAACGCCGTGCTGACCCACGTG AAGATCAACGACAAGTGCCCCAGAGAAGGCGAAGCCCACCTGGCCG AAGAGAACGAGGGCGACAACGCCTGCAAGCGGACCTACAGCGATAG AGGCAGAGGCAATGGCTGCGGCAGATTTGGCAAGGGCAGCATCGTG GCCTGCGCCAAGTTCACCTGTGCCAAGAGCATGAGCCTGTTCGAGG TGGACCAGACCAAGATCCAGTACGTGATCCGGGCCCAGCTGCACGT GGGAGCCAAGCAGGAAAACTGGAACACCGACATCAAGACCCTGAAG TTCGACGCCCTGAGCGGCTCCCAGGAAGTGGAATTCATCGGCTATG GCAAGGCCACCCTGGAATGTCAGGTGCAGACCGCCGTGGACTTCGG CAACAGCTATATCGCCGAGATGGAAACCGAGAGCTGGATCGTGGAC CGGCAGTGGGCTCAGGATCTGACCCTGCCTTGGCAGTCTGGCTCTG GCGGAGTGTGGCGGGAAATGCACCACCTGGTGGAATTCGAGCCTCC CCACGCCGCCACCATTAGAGTGCTGGCCCTGGGCAATCAGGAAGGC TCTCTGAAAACAGCCCTGACCGGCGCCATGAGAGTGACCAAGGACA CCAACGACAACAACCTGTACAAGCTGCATGGCGGCCACGTGTCCTG CAGAGTGAAGCTGTCTGCCCTGACACTGAAGGGCACCAGCTACAAG ATCTGCACCGACAAGATGTTCTTCGTGAAGAACCCCACCGACACCG GCCACGGCACAGTCGTGATGCAAGTGAAGGTGTCCAAGGGCGCTCC CTGCCGGATCCCTGTGATCGTGGCCGATGATCTGACAGCCGCCATC AACAAGGGCATCCTCGTGACAGTGAACCCTATCGCCTCCACCAACG ATGACGAGGTGCTGATCGAAGTGAATCCTCCCTTCGGCGACTCCTA CATCATCGTGGGACGGGGCGACAGCAGACTGACCTACCAGTGGCAC AAAGAGGGCAGCAGCATCGGCAAGCTGTTCACCCAGACCATGAAGG GCGTGGAACGGCTGGCCGTGATGGGAGATACCGCCTGGGATTTCTC TAGCGCTGGCGGCTTCTTCACCAGCGTGGGCAAGGGAATCCACACC GTGTTCGGCAGCGCCTTCCAGGGACTGTTCGGCGGCCTGAACTGGA TCACCAAAGTGATCATGGGCGCTGTGCTGATCTGGGTGGGAATCAA CACCCGGAACATGACCATGAGCATGTCCATGATCCTCGTGGGAGTG ATTATGATGTTCCTGAGCCTGGGTGTGGGCGCC YF_17D_JEVsp_ ATGTGGCTGGTGTCCCTGGCCATCGTGACAGCCTGTGCTGGCGCTG 78 SE_no_polyNs TGACACTCGTGCGGAAGAACAGATGGCTGCTGCTGAACGTGACCAG CGAGGACCTGGGCAAGACCTTCTCTGTAGGCACCGGCAACTGCACC ACCAACATCCTGGAAGCCAAGTACTGGTGCCCCGACAGCATGGAGT ACAACTGCCCCAACCTGAGCCCCAGAGAGGAACCCGACGACATCGA CTGCTGGTGCTACGGCGTGGAAAACGTGCGGGTGGCCTACGGCAAG TGCGATAGCGCCGGCAGAAGCAGAAGAAGCAGGCGGGCCATCGACC TGCCCACCCACGAAAACCACGGCCTGAAAACCCGGCAGGAAAAGTG GATGACCGGCCGGATGGGCGAGCGGCAGCTGCAGAAAATTGAGCGG TGGTTTGTGCGGAACCCCTTCTTCGCCGTGACCGCCCTGACAATCG CCTACCTCGTGGGCAGCAACATGACCCAGAGAGTCGTGATCGCCCT GCTGGTGCTGGCTGTGGGCCCTGCCTATAGCGCCATTGATCTGCCT ACACACGAGAATCATGGGCTGAAAACAAGACAGGAGAAATGGATGA CTGGGCGCATGGGAGAAAGACAGCTGCAGAAAATCGAACGGTGGTT CGTGCGCAATCCTTTCTTTGCTGTGACTGCTCTGACCATTGCCTAT CTCGTGGGATCCAATATGACACAGCGGGTCGTGATTGCTCTGCTGG TGCTGGCAGTGGGACCCGCTTACTCCGCCCACTGTATCGGCATCAC CGACCGGGACTTCATCGAGGGCGTGCACGGCGGAACATGGGTGTCC GCTACCCTGGAACAGGATAAGTGCGTGACCGTGATGGCCCCCGACA AGCCCAGCCTGGACATCAGCCTGGAAACCGTGGCCATCGATAGACC CGCCGAAGTGCGGAAAGTGTGCTACAACGCCGTGCTGACCCACGTG AAGATCAACGACAAGTGCCCCAGCACCGGCGAAGCCCACCTGGCCG AAGAGAACGAGGGCGACAACGCCTGCAAGCGGACCTACAGCGATAG AGGCTGGGGCAATGGCTGCGGCCTGTTTGGCAAGGGCAGCATCGTG GCCTGCGCCAAGTTCACCTGTGCCAAGAGCATGAGCCTGTTCGAGG TGGACCAGACCAAGATCCAGTACGTGATCCGGGCCCAGCTGCACGT GGGAGCCAAGCAGGAAAACTGGAACACCGACATCAAGACCCTGAAG TTCGACGCCCTGAGCGGCTCCCAGGAAGTGGAATTCATCGGCTATG GCAAGGCCACCCTGGAATGTCAGGTGCAGACCGCCGTGGACTTCGG CAACAGCTATATCGCCGAGATGGAAACCGAGAGCTGGATCGTGGAC CGGCAGTGGGCTCAGGATCTGACCCTGCCTTGGCAGTCTGGCTCTG GCGGAGTGTGGCGGGAAATGCACCACCTGGTGGAATTCGAGCCTCC CCACGCCGCCACCATTAGAGTGCTGGCCCTGGGCAATCAGGAAGGC TCTCTGAAAACAGCCCTGACCGGCGCCATGAGAGTGACCAAGGACA CCAACGACAACAACCTGTACAAGCTGCATGGCGGCCACGTGTCCTG CAGAGTGAAGCTGTCTGCCCTGACACTGAAGGGCACCAGCTACAAG ATCTGCACCGACAAGATGTTCTTCGTGAAGAACCCCACCGACACCG GCCACGGCACAGTCGTGATGCAAGTGAAGGTGTCCAAGGGCGCTCC CTGCCGGATCCCTGTGATCGTGGCCGATGATCTGACAGCCGCCATC AACAAGGGCATCCTCGTGACAGTGAACCCTATCGCCTCCACCAACG ATGACGAGGTGCTGATCGAAGTGAATCCTCCCTTCGGCGACTCCTA CATCATCGTGGGACGGGGCGACAGCAGACTGACCTACCAGTGGCAC AAAGAGGGCAGCAGCATCGGCAAGCTGTTCACCCAGACCATGAAGG GCGTGGAACGGCTGGCCGTGATGGGAGATACCGCCTGGGATTTCTC TAGCGCTGGCGGCTTCTTCACCAGCGTGGGCAAGGGAATCCACACC GTGTTCGGCAGCGCCTTCCAGGGACTGTTCGGCGGCCTGAACTGGA TCACCAAAGTGATCATGGGCGCTGTGCTGATCTGGGTGGGAATCAA CACCCGGAACATGACCATGAGCATGTCCATGATCCTCGTGGGAGTG ATTATGATGTTCCTGAGCCTGGGTGTGGGCGCC YF_17D_IgGksp_ ATGGAAACCCCTGCCCAGCTGCTGTTCCTGCTGCTGCTGTGGCTGC 79 SE_no_polyNs_ CTGATACCACCGGCGTGACACTCGTGCGGAAGAACAGATGGCTGCT mutFL GCTGAACGTGACCAGCGAGGACCTGGGCAAGACCTTCTCTGTGGGC ACCGGCAACTGCACCACCAACATCCTGGAAGCCAAGTACTGGTGCC CCGACAGCATGGAGTACAACTGCCCCAACCTGAGCCCCAGAGAGGA ACCCGACGACATCGACTGCTGGTGCTACGGCGTGGAAAACGTGCGG GTGGCCTACGGCAAGTGCGATAGCGCCGGCAGAAGCAGAAGAAGCA GGCGGGCCATCGACCTGCCCACCCACGAAAACCACGGCCTGAAAAC CCGGCAGGAAAAGTGGATGACCGGCCGGATGGGCGAGCGGCAGCTG CAGAAAATTGAGCGGTGGTTTGTGCGGAACCCCTTCTTCGCCGTGA CCGCCCTGACAATCGCCTACCTCGTGGGCAGCAACATGACCCAGAG AGTCGTGATCGCCCTGCTGGTGCTGGCTGTGGGCCCTGCCTATAGC GCCATTGATCTGCCTACACACGAGAATCATGGGCTGAAAACAAGAC AGGAGAAATGGATGACTGGGCGCATGGGAGAAAGACAGCTGCAGAA AATCGAACGGTGGTTCGTGCGCAATCCGTTCTTTGCTGTGACTGCT CTGACCATTGCCTATCTCGTGGGATCCAATATGACACAGCGGGTCG TGATTGCTCTGCTGGTGCTGGCAGTGGGACCCGCTTACTCCGCCCA CTGTATCGGCATCACCGACCGGGACTTCATCGAGGGCGTGCACGGC GGAACATGGGTGTCCGCTACCCTGGAACAGGATAAGTGCGTGACCG TGATGGCCCCCGACAAGCCCAGCCTGGACATCAGCCTGGAAACCGT GGCCATCGATAGACCCGCCGAAGTGCGGAAAGTGTGCTACAACGCC GTGCTGACCCACGTGAAGATCAACGACAAGTGCCCCAGAGAAGGCG AAGCCCACCTGGCCGAAGAGAACGAGGGCGACAACGCCTGCAAGCG GACCTACAGCGATAGAGGCAGAGGCAATGGCTGCGGCAGATTTGGC AAGGGCAGCATCGTGGCCTGCGCCAAGTTCACCTGTGCCAAGAGCA TGAGCCTGTTCGAGGTGGACCAGACCAAGATCCAGTACGTGATCCG GGCTCAGCTGCACGTGGGCGCCAAGCAGGAAAACTGGAACACCGAC ATCAAGACCCTGAAGTTCGACGCCCTGAGCGGCTCCCAGGAAGTGG AATTCATCGGCTATGGCAAGGCCACCCTGGAATGTCAGGTGCAGAC CGCCGTGGACTTCGGCAACAGCTATATCGCCGAGATGGAAACCGAG AGCTGGATCGTGGACCGGCAGTGGGCCCAGGATCTGACACTGCCTT GGCAGTCTGGCTCTGGCGGAGTGTGGCGGGAAATGCACCACCTGGT GGAATTCGAGCCTCCCCACGCCGCCACCATTAGAGTGCTGGCCCTG GGCAATCAGGAAGGCTCTCTGAAAACAGCCCTGACCGGCGCCATGA GAGTGACCAAGGACACCAACGACAACAACCTGTACAAGCTGCACGG GGGGCACGTGTCCTGCAGAGTGAAACTGTCTGCCCTGACACTGAAG GGCACCAGCTACAAGATCTGCACCGACAAGATGTTCTTCGTGAAGA ACCCCACCGACACCGGCCACGGCACAGTCGTGATGCAAGTGAAGGT GTCCAAGGGCGCTCCCTGCCGGATCCCTGTGATCGTGGCCGATGAT CTGACAGCCGCCATCAACAAGGGCATCCTCGTGACAGTGAACCCTA TCGCCTCCACCAACGATGACGAGGTGCTGATCGAAGTGAACCCTCC TTTCGGCGACTCCTACATCATCGTGGGACGGGGCGACAGCAGACTG ACCTACCAGTGGCACAAAGAGGGCAGCAGCATCGGCAAGCTGTTCA CCCAGACCATGAAGGGCGTGGAACGGCTGGCCGTGATGGGAGATAC CGCCTGGGATTTCTCTAGCGCTGGCGGCTTCTTCACCAGCGTGGGC AAGGGAATCCACACCGTGTTCGGCAGCGCCTTCCAGGGACTGTTCG GCGGCCTGAACTGGATCACCAAAGTGATCATGGGCGCTGTGCTGAT CTGGGTGGGAATCAACACCCGGAACATGACCATGAGCATGTCCATG ATCCTCGTGGGAGTGATTATGATGTTCCTGAGCCTGGGCGTGGGTG CC YF_17D_IgGksp_ ATGGAAACCCCTGCCCAGCTGCTGTTCCTGCTGCTGCTGTGGCTGC 80 SE_no_polyNs CTGATACCACCGGCGTGACACTCGTGCGGAAGAACAGATGGCTGCT GCTGAACGTGACCAGCGAGGACCTGGGCAAGACCTTCTCTGTGGGC ACCGGCAACTGCACCACCAACATCCTGGAAGCCAAGTACTGGTGCC CCGACAGCATGGAGTACAACTGCCCCAACCTGAGCCCCAGAGAGGA ACCCGACGACATCGACTGCTGGTGCTACGGCGTGGAAAACGTGCGG GTGGCCTACGGCAAGTGCGATAGCGCCGGCAGAAGCAGAAGAAGCA GGCGGGCCATCGACCTGCCCACCCACGAAAACCACGGCCTGAAAAC CCGGCAGGAAAAGTGGATGACCGGCCGGATGGGCGAGCGGCAGCTG CAGAAAATTGAGCGGTGGTTTGTGCGGAACCCCTTCTTCGCCGTGA CCGCCCTGACAATCGCCTACCTCGTGGGCAGCAACATGACCCAGAG AGTCGTGATCGCCCTGCTGGTGCTGGCTGTGGGCCCTGCCTATAGC GCCATTGATCTGCCTACACACGAGAATCATGGGCTGAAAACAAGAC AGGAGAAATGGATGACTGGGCGCATGGGAGAAAGACAGCTGCAGAA AATCGAACGGTGGTTCGTGCGCAATCCGTTCTTTGCTGTGACTGCT CTGACCATTGCCTATCTCGTGGGATCCAATATGACACAGCGGGTCG TGATTGCTCTGCTGGTGCTGGCAGTGGGACCCGCTTACTCCGCCCA CTGTATCGGCATCACCGACCGGGACTTCATCGAGGGCGTGCACGGC GGAACATGGGTGTCCGCTACCCTGGAACAGGATAAGTGCGTGACCG TGATGGCCCCCGACAAGCCCAGCCTGGACATCAGCCTGGAAACCGT GGCCATCGATAGACCCGCCGAAGTGCGGAAAGTGTGCTACAACGCC GTGCTGACCCACGTGAAGATCAACGACAAGTGCCCCAGCACCGGCG AAGCCCACCTGGCCGAAGAGAACGAGGGCGACAACGCCTGCAAGCG GACCTACAGCGATAGAGGCTGGGGCAATGGCTGCGGCCTGTTTGGC AAGGGCAGCATCGTGGCCTGCGCCAAGTTCACCTGTGCCAAGAGCA TGAGCCTGTTCGAGGTGGACCAGACCAAGATCCAGTACGTGATCCG GGCTCAGCTGCACGTGGGCGCCAAGCAGGAAAACTGGAACACCGAC ATCAAGACCCTGAAGTTCGACGCCCTGAGCGGCTCCCAGGAAGTGG AATTCATCGGCTATGGCAAGGCCACCCTGGAATGTCAGGTGCAGAC CGCCGTGGACTTCGGCAACAGCTATATCGCCGAGATGGAAACCGAG AGCTGGATCGTGGACCGGCAGTGGGCCCAGGATCTGACACTGCCTT GGCAGTCTGGCTCTGGCGGAGTGTGGCGGGAAATGCACCACCTGGT GGAATTCGAGCCTCCCCACGCCGCCACCATTAGAGTGCTGGCCCTG GGCAATCAGGAAGGCTCTCTGAAAACAGCCCTGACCGGCGCCATGA GAGTGACCAAGGACACCAACGACAACAACCTGTACAAGCTGCACGG GGGGCACGTGTCCTGCAGAGTGAAACTGTCTGCCCTGACACTGAAG GGCACCAGCTACAAGATCTGCACCGACAAGATGTTCTTCGTGAAGA ACCCCACCGACACCGGCCACGGCACAGTCGTGATGCAAGTGAAGGT GTCCAAGGGCGCTCCCTGCCGGATCCCTGTGATCGTGGCCGATGAT CTGACAGCCGCCATCAACAAGGGCATCCTCGTGACAGTGAACCCTA TCGCCTCCACCAACGATGACGAGGTGCTGATCGAAGTGAACCCTCC TTTCGGCGACTCCTACATCATCGTGGGACGGGGCGACAGCAGACTG ACCTACCAGTGGCACAAAGAGGGCAGCAGCATCGGCAAGCTGTTCA CCCAGACCATGAAGGGCGTGGAACGGCTGGCCGTGATGGGAGATAC CGCCTGGGATTTCTCTAGCGCTGGCGGCTTCTTCACCAGCGTGGGC AAGGGAATCCACACCGTGTTCGGCAGCGCCTTCCAGGGACTGTTCG GCGGCCTGAACTGGATCACCAAAGTGATCATGGGCGCTGTGCTGAT CTGGGTGGGAATCAACACCCGGAACATGACCATGAGCATGTCCATG ATCCTCGTGGGAGTGATTATGATGTTCCTGAGCCTGGGCGTGGGTG CC YFV mRNA Sequences YF_Con_prMe_ AUGGAAACCCCUGCCCAGCUGCUGUUCCUGCUGCUGCUGUGGCUGC 81 Sen2000_IgGksp CUGAUACCACCGGCGUGACACUCGUGCGGAAGAACAGAUGGCUGCU GCUGAACGUGACCAGCGAGGACCUGGGCAAGACCUUCUCUGUGGGC ACCGGCAACUGCACCACCAACAUCCUGGAAGCCAAGUACUGGUGCC CCGACAGCAUGGAGUACAACUGCCCCAACCUGAGCCCCAGAGAGGA ACCCGACGACAUCGACUGCUGGUGCUACGGCGUGGAAAACGUGCGG GUGGCCUACGGCAAGUGCGAUAGCGCCGGCAGAAGCAGAAGAAGCA GGCGGGCCAUCGACCUGCCCACCCACGAAAACCACGGCCUGAAAAC CCGGCAGGAAAAGUGGAUGACCGGCCGGAUGGGCGAGCGGCAGCUG CAGAAAAUCGAGAGAUGGCUCGUGCGCAACCCCUUCUUCGCCGUGA CCGCCCUGACAAUCGCCUACCUCGUGGGCAGCAACAUGACCCAGAG AGUCGUGAUCGCCCUGCUGGUGCUGGCUGUGGGCCCUGCCUAUAGC GCCCACUGUAUCGGCAUCACCGACCGGGACUUCAUCGAGGGCGUGC ACGGCGGAACAUGGGUGUCCGCUACCCUGGAACAGGAUAAGUGCGU GACCGUGAUGGCCCCCGACAAGCCCAGCCUGGACAUCAGCCUGGAA ACCGUGGCCAUUGACGGACCCGCCGAGGCCAGAAAAGUGUGCUACA ACGCCGUGCUGACCCACGUGAAGAUCAACGACAAGUGCCCCAGCAC CGGCGAAGCCCACCUGGCCGAAGAGAACGAGGGCGACAACGCCUGC AAGCGGACCUACAGCGAUAGAGGCUGGGGCAAUGGCUGCGGCCUGU UUGGCAAGGGCAGCAUCGUGGCCUGCGCCAAGUUCACCUGUGCCAA GAGCAUGAGCCUGUUCGAGGUGGACCAGACCAAGAUCCAGUACGUG AUCCGGGCUCAGCUGCACGUGGGCGCCAAGCAGGAAAACUGGAACA CCGACAUCAAGACCCUGAAGUUCGACGCCCUGAGCGGCUCCCAGGA AGCCGAGUUUACCGGCUAUGGCAAGGCCACCCUGGAAUGCCAGGUG CAGACCGCCGUGGACUUCGGCAACAGCUAUAUCGCCGAGAUGGAAA AAGAAAGCUGGAUCGUGGACCGGCAGUGGGCCCAGGAUCUGACACU GCCUUGGCAGUCUGGCUCUGGCGGAGUGUGGCGGGAAAUGCACCAC CUGGUGGAAUUCGAGCCUCCCCACGCCGCCACCAUUAGAGUGCUGG CCCUGGGCAAUCAGGAAGGCUCUCUGAAAACAGCCCUGACCGGCGC CAUGAGAGUGACCAAGGACACCAACGACAACAACCUGUACAAGCUG CACGGGGGGCACGUGUCCUGCAGAGUGAAACUGUCUGCCCUGACAC UGAAGGGCACCAGCUACAAGAUGUGCACCGACAAGAUGAGCUUCGU GAAGAACCCCACCGACACCGGCCACGGCACAGUCGUGAUGCAAGUG AAGGUGCCCAAGGGCGCUCCCUGCAGAAUCCCUGUGAUCGUGGCCG AUGAUCUGACAGCCGCCAUCAACAAGGGCAUCCUCGUGACAGUGAA CCCUAUCGCCUCCACCAACGAUGACGAGGUGCUGAUCGAAGUGAAC CCCCCCUUCGGCGACUCCUACAUCAUCGUGGGCACAGGCGACAGCA GACUGACCUACCAGUGGCACAAAGAGGGCAGCAGCAUCGGCAAGCU GUUCACCCAGACCAUGAAGGGCGCCGAGAGACUGGCUGUGAUGGGA GAUGCCGCCUGGGACUUUAGCAGCGCUGGCGGCUUCUUUACCAGCG UGGGCAAGGGAAUCCACACCGUGUUCGGCAGCGCCUUCCAGGGACU GUUUGGCGGCCUGAGCUGGAUCACCAAAGUGAUCAUGGGCGCUGUG CUGAUCUGGGUGGGAAUCAACACCCGGAACAUGACCAUGAGCAUGU CCAUGAUCCUCGUGGGAGUGAUUAUGAUGUUCCUGAGCCUGGGCGU GGGAGCC YF_Con_prME_ AUGUGGCUGGUGUCCCUGGCCAUCGUGACAGCCUGUGCUGGCGCUG 82 Sen2000_JEVsp UGACACUCGUGCGGAAGAACAGAUGGCUGCUGCUGAACGUGACCAG CGAGGACCUGGGCAAGACCUUCUCUGUGGGCACCGGCAACUGCACC ACCAACAUCCUGGAAGCCAAGUACUGGUGCCCCGACAGCAUGGAGU ACAACUGCCCCAACCUGAGCCCCAGAGAGGAACCCGACGACAUCGA CUGCUGGUGCUACGGCGUGGAAAACGUGCGGGUGGCCUACGGCAAG UGCGAUAGCGCCGGCAGAAGCAGAAGAAGCAGGCGGGCCAUCGACC UGCCCACCCACGAAAACCACGGCCUGAAAACCCGGCAGGAAAAGUG GAUGACCGGCCGGAUGGGCGAGCGGCAGCUGCAGAAAAUCGAGAGA UGGCUCGUGCGCAACCCCUUCUUCGCCGUGACCGCCCUGACAAUCG CCUACCUCGUGGGCAGCAACAUGACCCAGAGAGUCGUGAUCGCCCU GCUGGUGCUGGCUGUGGGCCCUGCCUAUAGCGCCCACUGUAUCGGC AUCACCGACCGGGACUUCAUCGAGGGCGUGCACGGCGGAACAUGGG UGUCCGCUACCCUGGAACAGGAUAAGUGCGUGACCGUGAUGGCCCC CGACAAGCCCAGCCUGGACAUCAGCCUGGAAACCGUGGCCAUUGAC GGACCCGCCGAGGCCAGAAAAGUGUGCUACAACGCCGUGCUGACCC ACGUGAAGAUCAACGACAAGUGCCCCAGCACCGGCGAAGCCCACCU GGCCGAAGAGAACGAGGGCGACAACGCCUGCAAGCGGACCUACAGC GAUAGAGGCUGGGGCAAUGGCUGCGGCCUGUUUGGCAAGGGCAGCA UCGUGGCCUGCGCCAAGUUCACCUGUGCCAAGAGCAUGAGCCUGUU CGAGGUGGACCAGACCAAGAUCCAGUACGUGAUCCGGGCCCAGCUG CACGUGGGAGCCAAGCAGGAAAACUGGAACACCGACAUCAAGACCC UGAAGUUCGACGCCCUGAGCGGCUCCCAGGAAGCCGAGUUUACCGG CUAUGGCAAGGCCACCCUGGAAUGCCAGGUGCAGACCGCCGUGGAC UUCGGCAACAGCUAUAUCGCCGAGAUGGAAAAAGAAAGCUGGAUCG UGGACCGGCAGUGGGCCCAGGACCUGACACUGCCUUGGCAGUCUGG AUCUGGCGGCGUGUGGCGGGAAAUGCACCACCUGGUGGAAUUCGAG CCUCCCCAUGCCGCCACCAUCAGAGUGCUGGCCCUGGGCAAUCAGG AAGGCUCUCUGAAAACAGCCCUGACCGGCGCCAUGAGAGUGACCAA GGACACCAACGACAACAACCUGUACAAGCUGCAUGGCGGCCACGUG UCCUGCAGAGUGAAGCUGUCUGCCCUGACCCUGAAAGGCACCAGCU ACAAGAUGUGCACCGACAAGAUGAGCUUCGUGAAGAACCCCACCGA CACCGGCCACGGCACAGUCGUGAUGCAAGUGAAGGUGCCCAAGGGC GCUCCCUGCAGAAUCCCUGUGAUCGUGGCCGAUGAUCUGACAGCCG CCAUCAACAAGGGCAUCCUCGUGACAGUGAACCCUAUCGCCUCCAC CAACGAUGACGAGGUGCUGAUCGAAGUGAACCCCCCCUUCGGCGAC UCCUACAUCAUCGUGGGCACAGGCGACAGCAGACUGACCUACCAGU GGCACAAAGAGGGCAGCAGCAUCGGCAAGCUGUUCACCCAGACCAU GAAGGGCGCCGAGAGACUGGCUGUGAUGGGAGAUGCCGCCUGGGAC UUUAGCAGCGCUGGCGGCUUCUUUACCAGCGUGGGCAAGGGAAUCC ACACCGUGUUCGGCAGCGCCUUCCAGGGACUGUUUGGCGGCCUGAG CUGGAUCACCAAAGUGAUCAUGGGCGCUGUGCUGAUCUGGGUGGGA AUCAACACCCGGAACAUGACCAUGAGCAUGUCCAUGAUCCUCGUGG GAGUGAUUAUGAUGUUCCUGAGCCUGGGCGUGGGCGCC YF_Con_prME_ AUGUGGCUGGUGUCCCUGGCCAUCGUGACAGCCUGUGCUGGCGCUG 83 Sen2000_JEVsp_ UGACACUCGUGCGGAAGAACAGAUGGCUGCUGCUGAACGUGACCAG N153T CGAGGACCUGGGCAAGACCUUCUCUGUGGGCACCGGCAACUGCACC ACCAACAUCCUGGAAGCCAAGUACUGGUGCCCCGACAGCAUGGAGU ACAACUGCCCCAACCUGAGCCCCAGAGAGGAACCCGACGACAUCGA CUGCUGGUGCUACGGCGUGGAAAACGUGCGGGUGGCCUACGGCAAG UGCGAUAGCGCCGGCAGAAGCAGAAGAAGCAGGCGGGCCAUCGACC UGCCCACCCACGAAAACCACGGCCUGAAAACCCGGCAGGAAAAGUG GAUGACCGGCCGGAUGGGCGAGCGGCAGCUGCAGAAAAUCGAGAGA UGGCUCGUGCGCAACCCCUUCUUCGCCGUGACCGCCCUGACAAUCG CCUACCUCGUGGGCAGCAACAUGACCCAGAGAGUCGUGAUCGCCCU GCUGGUGCUGGCUGUGGGCCCUGCCUAUAGCGCCCACUGUAUCGGC AUCACCGACCGGGACUUCAUCGAGGGCGUGCACGGCGGAACAUGGG UGUCCGCUACCCUGGAACAGGAUAAGUGCGUGACCGUGAUGGCCCC CGACAAGCCCAGCCUGGACAUCAGCCUGGAAACCGUGGCCAUUGAC GGACCCGCCGAGGCCAGAAAAGUGUGCUACAACGCCGUGCUGACCC ACGUGAAGAUCAACGACAAGUGCCCCAGCACCGGCGAAGCCCACCU GGCCGAAGAGAACGAGGGCGACAACGCCUGCAAGCGGACCUACAGC GAUAGAGGCUGGGGCAAUGGCUGCGGCCUGUUUGGCAAGGGCAGCA UCGUGGCCUGCGCCAAGUUCACCUGUGCCAAGAGCAUGAGCCUGUU CGAGGUGGACCAGACCAAGAUCCAGUACGUGAUCCGGGCCCAGCUG CACGUGGGAGCCAAGCAGGAAAACUGGACCACCGACAUCAAGACCC UGAAGUUCGACGCCCUGAGCGGCUCCCAGGAAGCCGAGUUUACCGG CUAUGGCAAGGCCACCCUGGAAUGCCAGGUGCAGACCGCCGUGGAC UUCGGCAACAGCUAUAUCGCCGAGAUGGAAAAAGAAAGCUGGAUCG UGGACCGGCAGUGGGCCCAGGACCUGACACUGCCUUGGCAGUCUGG AUCUGGCGGCGUGUGGCGGGAAAUGCACCACCUGGUGGAAUUCGAG CCUCCCCAUGCCGCCACCAUCAGAGUGCUGGCCCUGGGCAAUCAGG AAGGCUCUCUGAAAACAGCCCUGACCGGCGCCAUGAGAGUGACCAA GGACACCAACGACAACAACCUGUACAAGCUGCAUGGCGGCCACGUG UCCUGCAGAGUGAAGCUGUCUGCCCUGACCCUGAAAGGCACCAGCU ACAAGAUGUGCACCGACAAGAUGAGCUUCGUGAAGAACCCCACCGA CACCGGCCACGGCACAGUCGUGAUGCAAGUGAAGGUGCCCAAGGGC GCUCCCUGCAGAAUCCCUGUGAUCGUGGCCGAUGAUCUGACAGCCG CCAUCAACAAGGGCAUCCUCGUGACAGUGAACCCUAUCGCCUCCAC CAACGAUGACGAGGUGCUGAUCGAAGUGAACCCCCCCUUCGGCGAC UCCUACAUCAUCGUGGGCACAGGCGACAGCAGACUGACCUACCAGU GGCACAAAGAGGGCAGCAGCAUCGGCAAGCUGUUCACCCAGACCAU GAAGGGCGCCGAGAGACUGGCUGUGAUGGGAGAUGCCGCCUGGGAC UUUAGCAGCGCUGGCGGCUUCUUUACCAGCGUGGGCAAGGGAAUCC ACACCGUGUUCGGCAGCGCCUUCCAGGGACUGUUUGGCGGCCUGAG CUGGAUCACCAAAGUGAUCAUGGGCGCUGUGCUGAUCUGGGUGGGA AUCAACACCCGGAACAUGACCAUGAGCAUGUCCAUGAUCCUCGUGG GAGUGAUUAUGAUGUUCCUGAGCCUGGGCGUGGGCGCC 17D_vaccine_ AUGUGGCUGGUGUCCCUGGCCAUCGUGACAGCCUGUGCUGGCGCUG 84 prME_JEVsp UGACACUCGUGCGGAAAAACAGAUGGCUGCUGCUGAACGUGACCAG CGAGGACCUGGGCAAGACCUUCUCUGUAGGCACCGGCAACUGCACC ACCAACAUCCUGGAAGCCAAGUACUGGUGCCCCGACAGCAUGGAGU ACAACUGCCCCAACCUGAGCCCCAGAGAGGAACCCGACGACAUCGA CUGCUGGUGCUACGGCGUGGAAAACGUGCGGGUGGCCUACGGCAAG UGCGAUAGCGCCGGCAGAAGCAGAAGAAGCAGGCGGGCCAUCGACC UGCCCACCCACGAAAACCACGGCCUGAAAACCCGGCAGGAAAAGUG GAUGACCGGCCGGAUGGGCGAGCGGCAGCUGCAGAAAAUUGAGCGG UGGUUUGUGCGGAACCCCUUCUUCGCCGUGACCGCCCUGACAAUCG CCUACCUCGUGGGCAGCAACAUGACCCAGAGAGUCGUGAUCGCCCU GCUGGUGCUGGCUGUGGGCCCUGCCUAUAGCGCCAUUGAUCUGCCU ACACACGAGAAUCAUGGGCUGAAAACAAGACAGGAAAAAUGGAUGA CUGGGCGCAUGGGAGAAAGACAGCUGCAGAAAAUCGAACGGUGGUU CGUGCGCAAUCCUUUUUUUGCUGUGACUGCUCUGACCAUUGCCUAU CUCGUGGGAUCCAAUAUGACACAGCGGGUCGUGAUUGCUCUGCUGG UGCUGGCAGUGGGACCCGCUUACUCCGCCCACUGUAUCGGCAUCAC CGACCGGGACUUCAUCGAGGGCGUGCACGGCGGAACAUGGGUGUCC GCUACCCUGGAACAGGAUAAGUGCGUGACCGUGAUGGCCCCCGACA AGCCCAGCCUGGACAUCAGCCUGGAAACCGUGGCCAUCGAUAGACC CGCCGAAGUGCGGAAAGUGUGCUACAACGCCGUGCUGACCCACGUG AAGAUCAACGACAAGUGCCCCAGCACCGGCGAAGCCCACCUGGCCG AAGAGAACGAGGGCGACAACGCCUGCAAGCGGACCUACAGCGAUAG AGGCUGGGGCAAUGGCUGCGGCCUGUUUGGCAAGGGCAGCAUCGUG GCCUGCGCCAAGUUCACCUGUGCCAAGAGCAUGAGCCUGUUCGAGG UGGACCAGACCAAGAUCCAGUACGUGAUCCGGGCCCAGCUGCACGU GGGAGCCAAGCAGGAAAACUGGAACACCGACAUCAAGACCCUGAAG UUCGACGCCCUGAGCGGCUCCCAGGAAGUGGAAUUCAUCGGCUAUG GCAAGGCCACCCUGGAAUGUCAGGUGCAGACCGCCGUGGACUUCGG CAACAGCUAUAUCGCCGAGAUGGAAACCGAGAGCUGGAUCGUGGAC CGGCAGUGGGCUCAGGAUCUGACCCUGCCUUGGCAGUCUGGCUCUG GCGGAGUGUGGCGGGAAAUGCACCACCUGGUGGAAUUCGAGCCUCC CCACGCCGCCACCAUUAGAGUGCUGGCCCUGGGCAAUCAGGAAGGC UCUCUGAAAACAGCCCUGACCGGCGCCAUGAGAGUGACCAAGGACA CCAACGACAACAACCUGUACAAGCUGCAUGGCGGCCACGUGUCCUG CAGAGUGAAGCUGUCUGCCCUGACACUGAAGGGCACCAGCUACAAG AUCUGCACCGACAAGAUGUUCUUCGUGAAGAACCCCACCGACACCG GCCACGGCACAGUCGUGAUGCAAGUGAAGGUGUCCAAGGGCGCUCC CUGCCGGAUCCCUGUGAUCGUGGCCGAUGAUCUGACAGCCGCCAUC AACAAGGGCAUCCUCGUGACAGUGAACCCUAUCGCCUCCACCAACG AUGACGAGGUGCUGAUCGAAGUGAACCCCCCCUUCGGCGACUCCUA CAUCAUCGUGGGACGGGGCGACAGCAGACUGACCUACCAGUGGCAC AAAGAGGGCAGCAGCAUCGGCAAGCUGUUCACCCAGACCAUGAAGG GCGUGGAACGGCUGGCCGUGAUGGGAGAUACCGCCUGGGAUUUCUC UAGCGCUGGCGGCUUCUUCACCAGCGUGGGCAAGGGAAUCCACACC GUGUUCGGCAGCGCCUUCCAGGGACUGUUCGGCGGCCUGAACUGGA UCACCAAAGUGAUCAUGGGCGCUGUGCUGAUCUGGGUGGGAAUCAA CACCCGGAACAUGACCAUGAGCAUGUCCAUGAUCCUCGUGGGAGUG AUUAUGAUGUUCCUGAGCCUGGGUGUGGGCGCC 17D_vaccine_ AUGGAAACCCCUGCCCAGCUGCUGUUCCUGCUGCUGCUGUGGCUGC 85 prME_IgGKsp CUGAUACCACCGGCGUGACACUCGUGCGGAAGAACAGAUGGCUGCU GCUGAACGUGACCAGCGAGGACCUGGGCAAGACCUUCUCUGUGGGC ACCGGCAACUGCACCACCAACAUCCUGGAAGCCAAGUACUGGUGCC CCGACAGCAUGGAGUACAACUGCCCCAACCUGAGCCCCAGAGAGGA ACCCGACGACAUCGACUGCUGGUGCUACGGCGUGGAAAACGUGCGG GUGGCCUACGGCAAGUGCGAUAGCGCCGGCAGAAGCAGAAGAAGCA GGCGGGCCAUCGACCUGCCCACCCACGAAAACCACGGCCUGAAAAC CCGGCAGGAAAAGUGGAUGACCGGCCGGAUGGGCGAGCGGCAGCUG CAGAAAAUUGAGCGGUGGUUUGUGCGGAACCCCUUCUUCGCCGUGA CCGCCCUGACAAUCGCCUACCUCGUGGGCAGCAACAUGACCCAGAG AGUCGUGAUCGCCCUGCUGGUGCUGGCUGUGGGCCCUGCCUAUAGC GCCAUUGAUCUGCCUACACACGAGAAUCAUGGGCUGAAAACAAGAC AGGAAAAAUGGAUGACUGGGCGCAUGGGAGAAAGACAGCUGCAGAA AAUCGAACGGUGGUUCGUGCGCAAUCCUUUUUUUGCUGUGACUGCU CUGACCAUUGCCUAUCUCGUGGGAUCCAAUAUGACACAGCGGGUCG UGAUUGCUCUGCUGGUGCUGGCAGUGGGACCCGCUUACUCCGCCCA CUGUAUCGGCAUCACCGACCGGGACUUCAUCGAGGGCGUGCACGGC GGAACAUGGGUGUCCGCUACCCUGGAACAGGAUAAGUGCGUGACCG UGAUGGCCCCCGACAAGCCCAGCCUGGACAUCAGCCUGGAAACCGU GGCCAUCGAUAGACCCGCCGAAGUGCGGAAAGUGUGCUACAACGCC GUGCUGACCCACGUGAAGAUCAACGACAAGUGCCCCAGCACCGGCG AAGCCCACCUGGCCGAAGAGAACGAGGGCGACAACGCCUGCAAGCG GACCUACAGCGAUAGAGGCUGGGGCAAUGGCUGCGGCCUGUUUGGC AAGGGCAGCAUCGUGGCCUGCGCCAAGUUCACCUGUGCCAAGAGCA UGAGCCUGUUCGAGGUGGACCAGACCAAGAUCCAGUACGUGAUCCG GGCUCAGCUGCACGUGGGCGCCAAGCAGGAAAACUGGAACACCGAC AUCAAGACCCUGAAGUUCGACGCCCUGAGCGGCUCCCAGGAAGUGG AAUUCAUCGGCUAUGGCAAGGCCACCCUGGAAUGUCAGGUGCAGAC CGCCGUGGACUUCGGCAACAGCUAUAUCGCCGAGAUGGAAACCGAG AGCUGGAUCGUGGACCGGCAGUGGGCCCAGGAUCUGACACUGCCUU GGAAUUCGAGCCUCCCCACGCCGCCACCAUUAGAGUGCUGGCCCUG GGCAAUCAGGAAGGCUCUCUGAAAACAGCCCUGACCGGCGCCAUGA GAGUGACCAAGGACACCAACGACAACAACCUGUACAAGCUGCACGG GGGGCACGUGUCCUGCAGAGUGAAACUGUCUGCCCUGACACUGAAG GGCACCAGCUACAAGAUCUGCACCGACAAGAUGUUCUUCGUGAAGA ACCCCACCGACACCGGCCACGGCACAGUCGUGAUGCAAGUGAAGGU GUCCAAGGGCGCUCCCUGCCGGAUCCCUGUGAUCGUGGCCGAUGAU CUGACAGCCGCCAUCAACAAGGGCAUCCUCGUGACAGUGAACCCUA UCGCCUCCACCAACGAUGACGAGGUGCUGAUCGAAGUGAACCCCCC CUUCGGCGACUCCUACAUCAUCGUGGGACGGGGCGACAGCAGACUG ACCUACCAGUGGCACAAAGAGGGCAGCAGCAUCGGCAAGCUGUUCA CCCAGACCAUGAAGGGCGUGGAACGGCUGGCCGUGAUGGGAGAUAC CGCCUGGGAUUUCUCUAGCGCUGGCGGCUUCUUCACCAGCGUGGGC AAGGGAAUCCACACCGUGUUCGGCAGCGCCUUCCAGGGACUGUUCG GCGGCCUGAACUGGAUCACCAAAGUGAUCAUGGGCGCUGUGCUGAU CUGGGUGGGAAUCAACACCCGGAACAUGACCAUGAGCAUGUCCAUG AUCCUCGUGGGAGUGAUUAUGAUGUUCCUGAGCCUGGGCGUGGGUG CC YF_Sen2000_ AUGUGGCUGGUGUCCCUGGCCAUCGUGACAGCCUGUGCUGGCGCUG 86 JEVsp_N153T UGACACUCGUGCGGAAGAACAGAUGGCUGCUGCUGAACGUGACCAG CGAGGACCUGGGCAAGACCUUCUCUGUGGGCACCGGCAACUGCACC ACCAACAUCCUGGAAGCCAAGUACUGGUGCCCCGACAGCAUGGAGU ACAACUGCCCCAACCUGAGCCCCAGAGAGGAACCCGACGACAUCGA CUGCUGGUGCUACGGCGUGGAAAACGUGCGGGUGGCCUACGGCAAG UGCGAUAGCGCCGGCAGAAGCAGAAGAAGCAGGCGGGCCAUCGACC UGCCCACCCACGAAAACCACGGCCUGAAAACCCGGCAGGAAAAGUG GAUGACCGGCCGGAUGGGCGAGCGGCAGCUGCAGAAAAUCGAGAGA UGGCUCGUGCGCAACCCCUUCUUCGCCGUGACCGCCCUGACAAUCG CCUACCUCGUGGGCAGCAACAUGACCCAGAGAGUCGUGAUCGCCCU GCUGGUGCUGGCUGUGGGCCCUGCCUAUAGCGCCCACUGUAUCGGC AUCACCGACCGGGACUUCAUCGAGGGCGUGCACGGCGGAACAUGGG UGUCCGCUACCCUGGAACAGGAUAAGUGCGUGACCGUGAUGGCCCC CGACAAGCCCAGCCUGGACAUCAGCCUGGAAACCGUGGCCAUUGAC GGACCCGCCGAGGCCAGAAAAGUGUGCUACAACGCCGUGCUGACCC ACGUGAAGAUCAACGACAAGUGCCCCAGCACCGGCGAAGCCCACCU GGCCGAAGAGAACGAGGGCGACAACGCCUGCAAGCGGACCUACAGC GAUAGAGGCUGGGGCAAUGGCUGCGGCCUGUUUGGCAAGGGCAGCA UCGUGGCCUGCGCCAAGUUCACCUGUGCCAAGAGCAUGAGCCUGUU CGAGGUGGACCAGACCAAGAUCCAGUACGUGAUCCGGGCCCAGCUG CACGUGGGAGCCAAGCAGGAAAACUGGACCACCGACAUCAAGACCC UGAAGUUCGACGCCCUGAGCGGCUCCCAGGAAGCCGAGUUUACCGG CUAUGGCAAGGCCACCCUGGAAUGCCAGGUGCAGACCGCCGUGGAC UUCGGCAACAGCUAUAUCGCCGAGAUGGAAAAAGAAAGCUGGAUCG UGGACCGGCAGUGGGCCCAGGACCUGACACUGCCUUGGCAGUCUGG AUCUGGCGGCGUGUGGCGGGAAAUGCACCACCUGGUGGAAUUCGAG CCUCCCCAUGCCGCCACCAUCAGAGUGCUGGCCCUGGGCAAUCAGG AAGGCUCUCUGAAAACAGCCCUGACCGGCGCCAUGAGAGUGACCAA GGACACCAACGACAACAACCUGUACAAGCUGCAUGGCGGCCACGUG UCCUGCAGAGUGAAGCUGUCUGCCCUGACCCUGAAAGGCACCAGCU ACAAGAUGUGCACCGACAAGAUGAGCUUCGUGAAGAACCCCACCGA CACCGGCCACGGCACAGUCGUGAUGCAAGUGAAGGUGCCCAAGGGC GCUCCCUGCAGAAUCCCUGUGAUCGUGGCCGAUGAUCUGACAGCCG CCAUCAACAAGGGCAUCCUCGUGACAGUGAACCCUAUCGCCUCCAC CAACGAUGACGAGGUGCUGAUCGAAGUGAACCCCCCCUUCGGCGAC UCCUACAUCAUCGUGGGCACAGGCGACAGCAGACUGACCUACCAGU GGCACAAAGAGGGCAGCAGCAUCGGCAAGCUGUUCACCCAGACCAU GAAGGGCGCCGAGAGACUGGCUGUGAUGGGAGAUGCCGCCUGGGAC UUUAGCAGCGCUGGCGGCUUCUUUACCAGCGUGGGCAAGGGAAUCC ACACCGUGUUCGGCAGCGCCUUCCAGGGACUGUUUGGCGGCCUGAG CUGGAUCACCAAAGUGAUCAUGGGCGCUGUGCUGAUCUGGGUGGGA AUCAACACCCGGAACAUGACCAUGAGCAUGUCCAUGAUCCUCGUGG GAGUGAUUAUGAUGUUCCUGAGCCUGGGCGUGGGCGCC YF_17D_IgGksp_SE AUGGAAACCCCUGCCCAGCUGCUGUUCCUGCUGCUGCUGUGGCUGC 87 CUGAUACCACCGGCGUGACACUCGUGCGGAAGAACAGAUGGCUGCU GCUGAACGUGACCAGCGAGGACCUGGGCAAGACCUUCUCUGUGGGC ACCGGCAACUGCACCACCAACAUCCUGGAAGCCAAGUACUGGUGCC CCGACAGCAUGGAGUACAACUGCCCCAACCUGAGCCCCAGAGAGGA ACCCGACGACAUCGACUGCUGGUGCUACGGCGUGGAAAACGUGCGG GUGGCCUACGGCAAGUGCGAUAGCGCCGGCAGAAGCAGAAGAAGCA GGCGGGCCAUCGACCUGCCCACCCACGAAAACCACGGCCUGAAAAC CCGGCAGGAAAAGUGGAUGACCGGCCGGAUGGGCGAGCGGCAGCUG CAGAAAAUUGAGCGGUGGUUUGUGCGGAACCCCUUCUUCGCCGUGA CCGCCCUGACAAUCGCCUACCUCGUGGGCAGCAACAUGACCCAGAG AGUCGUGAUCGCCCUGCUGGUGCUGGCUGUGGGCCCUGCCUAUAGC GCCAUUGAUCUGCCUACACACGAGAAUCAUGGGCUGAAAACAAGAC AGGAAAAAUGGAUGACUGGGCGCAUGGGAGAAAGACAGCUGCAGAA AAUCGAACGGUGGUUCGUGCGCAAUCCUUUUUUUGCUGUGACUGCU CUGACCAUUGCCUAUCUCGUGGGAUCCAAUAUGACACAGCGGGUCG UGAUUGCUCUGCUGGUGCUGGCAGUGGGACCCGCUUACUCCGCCCA CUGUAUCGGCAUCACCGACCGGGACUUCAUCGAGGGCGUGCACGGC GGAACAUGGGUGUCCGCUACCCUGGAACAGGAUAAGUGCGUGACCG UGAUGGCCCCCGACAAGCCCAGCCUGGACAUCAGCCUGGAAACCGU GGCCAUCGAUAGACCCGCCGAAGUGCGGAAAGUGUGCUACAACGCC GUGCUGACCCACGUGAAGAUCAACGACAAGUGCCCCAGCACCGGCG AAGCCCACCUGGCCGAAGAGAACGAGGGCGACAACGCCUGCAAGCG GACCUACAGCGAUAGAGGCUGGGGCAAUGGCUGCGGCCUGUUUGGC AAGGGCAGCAUCGUGGCCUGCGCCAAGUUCACCUGUGCCAAGAGCA UGAGCCUGUUCGAGGUGGACCAGACCAAGAUCCAGUACGUGAUCCG GGCUCAGCUGCACGUGGGCGCCAAGCAGGAAAACUGGAACACCGAC AUCAAGACCCUGAAGUUCGACGCCCUGAGCGGCUCCCAGGAAGUGG AAUUCAUCGGCUAUGGCAAGGCCACCCUGGAAUGUCAGGUGCAGAC CGCCGUGGACUUCGGCAACAGCUAUAUCGCCGAGAUGGAAACCGAG AGCUGGAUCGUGGACCGGCAGUGGGCCCAGGAUCUGACACUGCCUU GGCAGUCUGGCUCUGGCGGAGUGUGGCGGGAAAUGCACCACCUGGU GGAAUUCGAGCCUCCCCACGCCGCCACCAUUAGAGUGCUGGCCCUG GGCAAUCAGGAAGGCUCUCUGAAAACAGCCCUGACCGGCGCCAUGA GAGUGACCAAGGACACCAACGACAACAACCUGUACAAGCUGCACGG GGGGCACGUGUCCUGCAGAGUGAAACUGUCUGCCCUGACACUGAAG GGCACCAGCUACAAGAUCUGCACCGACAAGAUGUUCUUCGUGAAGA ACCCCACCGACACCGGCCACGGCACAGUCGUGAUGCAAGUGAAGGU GUCCAAGGGCGCUCCCUGCCGGAUCCCUGUGAUCGUGGCCGAUGAU CUGACAGCCGCCAUCAACAAGGGCAUCCUCGUGACAGUGAACCCUA UCGCCUCCACCAACGAUGACGAGGUGCUGAUCGAAGUGAACCCCCC CUUCGGCGACUCCUACAUCAUCGUGGGACGGGGCGACAGCAGACUG ACCUACCAGUGGCACAAAGAGGGCAGCAGCAUCGGCAAGCUGUUCA CCCAGACCAUGAAGGGCGUGGAACGGCUGGCCGUGAUGGGAGAUAC CGCCUGGGAUUUCUCUAGCGCUGGCGGCUUCUUCACCAGCGUGGGC AAGGGAAUCCACACCGUGUUCGGCAGCGCCUUCCAGGGACUGUUCG GCGGCCUGAACUGGAUCACCAAAGUGAUCAUGGGCGCUGUGCUGAU CUGGGUGGGAAUCAACACCCGGAACAUGACCAUGAGCAUGUCCAUG AUCCUCGUGGGAGUGAUUAUGAUGUUCCUGAGCCUGGGCGUGGGUG CC YF_17D_JEVsp_SE AUGUGGCUGGUGUCCCUGGCCAUCGUGACAGCCUGUGCUGGCGCUG 88 UGACACUCGUGCGGAAAAACAGAUGGCUGCUGCUGAACGUGACCAG CGAGGACCUGGGCAAGACCUUCUCUGUAGGCACCGGCAACUGCACC ACCAACAUCCUGGAAGCCAAGUACUGGUGCCCCGACAGCAUGGAGU ACAACUGCCCCAACCUGAGCCCCAGAGAGGAACCCGACGACAUCGA CUGCUGGUGCUACGGCGUGGAAAACGUGCGGGUGGCCUACGGCAAG UGCGAUAGCGCCGGCAGAAGCAGAAGAAGCAGGCGGGCCAUCGACC UGCCCACCCACGAAAACCACGGCCUGAAAACCCGGCAGGAAAAGUG GAUGACCGGCCGGAUGGGCGAGCGGCAGCUGCAGAAAAUUGAGCGG UGGUUUGUGCGGAACCCCUUCUUCGCCGUGACCGCCCUGACAAUCG CCUACCUCGUGGGCAGCAACAUGACCCAGAGAGUCGUGAUCGCCCU GCUGGUGCUGGCUGUGGGCCCUGCCUAUAGCGCCAUUGAUCUGCCU ACACACGAGAAUCAUGGGCUGAAAACAAGACAGGAAAAAUGGAUGA CUGGGCGCAUGGGAGAAAGACAGCUGCAGAAAAUCGAACGGUGGUU CGUGCGCAAUCCUUUUUUUGCUGUGACUGCUCUGACCAUUGCCUAU CUCGUGGGAUCCAAUAUGACACAGCGGGUCGUGAUUGCUCUGCUGG UGCUGGCAGUGGGACCCGCUUACUCCGCCCACUGUAUCGGCAUCAC CGACCGGGACUUCAUCGAGGGCGUGCACGGCGGAACAUGGGUGUCC GCUACCCUGGAACAGGAUAAGUGCGUGACCGUGAUGGCCCCCGACA AGCCCAGCCUGGACAUCAGCCUGGAAACCGUGGCCAUCGAUAGACC CGCCGAAGUGCGGAAAGUGUGCUACAACGCCGUGCUGACCCACGUG AAGAUCAACGACAAGUGCCCCAGCACCGGCGAAGCCCACCUGGCCG AAGAGAACGAGGGCGACAACGCCUGCAAGCGGACCUACAGCGAUAG AGGCUGGGGCAAUGGCUGCGGCCUGUUUGGCAAGGGCAGCAUCGUG GCCUGCGCCAAGUUCACCUGUGCCAAGAGCAUGAGCCUGUUCGAGG UGGACCAGACCAAGAUCCAGUACGUGAUCCGGGCCCAGCUGCACGU GGGAGCCAAGCAGGAAAACUGGAACACCGACAUCAAGACCCUGAAG UUCGACGCCCUGAGCGGCUCCCAGGAAGUGGAAUUCAUCGGCUAUG GCAAGGCCACCCUGGAAUGUCAGGUGCAGACCGCCGUGGACUUCGG CAACAGCUAUAUCGCCGAGAUGGAAACCGAGAGCUGGAUCGUGGAC CGGCAGUGGGCUCAGGAUCUGACCCUGCCUUGGCAGUCUGGCUCUG GCGGAGUGUGGCGGGAAAUGCACCACCUGGUGGAAUUCGAGCCUCC CCACGCCGCCACCAUUAGAGUGCUGGCCCUGGGCAAUCAGGAAGGC UCUCUGAAAACAGCCCUGACCGGCGCCAUGAGAGUGACCAAGGACA CCAACGACAACAACCUGUACAAGCUGCAUGGCGGCCACGUGUCCUG CAGAGUGAAGCUGUCUGCCCUGACACUGAAGGGCACCAGCUACAAG AUCUGCACCGACAAGAUGUUCUUCGUGAAGAACCCCACCGACACCG GCCACGGCACAGUCGUGAUGCAAGUGAAGGUGUCCAAGGGCGCUCC CUGCCGGAUCCCUGUGAUCGUGGCCGAUGAUCUGACAGCCGCCAUC AACAAGGGCAUCCUCGUGACAGUGAACCCUAUCGCCUCCACCAACG AUGACGAGGUGCUGAUCGAAGUGAACCCCCCCUUCGGCGACUCCUA CAUCAUCGUGGGACGGGGCGACAGCAGACUGACCUACCAGUGGCAC AAAGAGGGCAGCAGCAUCGGCAAGCUGUUCACCCAGACCAUGAAGG GCGUGGAACGGCUGGCCGUGAUGGGAGAUACCGCCUGGGAUUUCUC UAGCGCUGGCGGCUUCUUCACCAGCGUGGGCAAGGGAAUCCACACC GUGUUCGGCAGCGCCUUCCAGGGACUGUUCGGCGGCCUGAACUGGA UCACCAAAGUGAUCAUGGGCGCUGUGCUGAUCUGGGUGGGAAUCAA CACCCGGAACAUGACCAUGAGCAUGUCCAUGAUCCUCGUGGGAGUG AUUAUGAUGUUCCUGAGCCUGGGUGUGGGCGCC YF_SEN2000_ AUGUGGCUGGUGUCCCUGGCCAUCGUGACAGCCUGUGCUGGCGCUG 89 prME_JEVsp UGACACUCGUGCGGAAGAACAGAUGGCUGCUGCUGAACGUGACCAG CGAGGACCUGGGCAAGACCUUCUCUGUGGGCACCGGCAACUGCACC ACCAACAUCCUGGAAGCCAAGUACUGGUGCCCCGACAGCAUGGAGU ACAACUGCCCCAACCUGAGCCCCAGAGAGGAACCCGACGACAUCGA CUGCUGGUGCUACGGCGUGGAAAACGUGCGGGUGGCCUACGGCAAG UGCGAUAGCGCCGGCAGAAGCAGAAGAAGCAGGCGGGCCAUCGACC UGCCCACCCACGAAAACCACGGCCUGAAAACCCGGCAGGAAAAGUG GAUGACCGGCCGGAUGGGCGAGCGGCAGCUGCAGAAAAUCGAGAGA UGGCUCGUGCGCAACCCCUUCUUCGCCGUGACCGCCCUGACAAUCG CCUACCUCGUGGGCAGCAACAUGACCCAGAGAGUCGUGAUCGCCCU GCUGGUGCUGGCUGUGGGCCCUGCCUAUAGCGCCCACUGUAUCGGC AUCACCGACCGGGACUUCAUCGAGGGCGUGCACGGCGGAACAUGGG UGUCCGCUACCCUGGAACAGGAUAAGUGCGUGACCGUGAUGGCCCC CGACAAGCCCAGCCUGGACAUCAGCCUGGAAACCGUGGCCAUUGAC GGACCCGCCGAGGCCAGAAAAGUGUGCUACAACGCCGUGCUGACCC ACGUGAAGAUCAACGACAAGUGCCCCAGCACCGGCGAAGCCCACCU GGCCGAAGAGAACGAGGGCGACAACGCCUGCAAGCGGACCUACAGC GAUAGAGGCUGGGGCAAUGGCUGCGGCCUGUUUGGCAAGGGCAGCA UCGUGGCCUGCGCCAAGUUCACCUGUGCCAAGAGCAUGAGCCUGUU CGAGGUGGACCAGACCAAGAUCCAGUACGUGAUCCGGGCCCAGCUG CACGUGGGAGCCAAGCAGGAAAACUGGAACACCGACAUCAAGACCC UGAAGUUCGACGCCCUGAGCGGCUCCCAGGAAGCCGAGUUUACCGG CUAUGGCAAGGCCACCCUGGAAUGCCAGGUGCAGACCGCCGUGGAC UUCGGCAACAGCUAUAUCGCCGAGAUGGAAAAAGAAAGCUGGAUCG UGGACCGGCAGUGGGCCCAGGACCUGACACUGCCUUGGCAGUCUGG AUCUGGCGGCGUGUGGCGGGAAAUGCACCACCUGGUGGAAUUCGAG CCUCCCCAUGCCGCCACCAUCAGAGUGCUGGCCCUGGGCAAUCAGG AAGGCUCUCUGAAAACAGCCCUGACCGGCGCCAUGAGAGUGACCAA GGACACCAACGACAACAACCUGUACAAGCUGCAUGGCGGCCACGUG UCCUGCAGAGUGAAGCUGUCUGCCCUGACCCUGAAAGGCACCAGCU ACAAGAUGUGCACCGACAAGAUGAGCUUCGUGAAGAACCCCACCGA CACCGGCCACGGCACAGUCGUGAUGCAAGUGAAGGUGCCCAAGGGC GCUCCCUGCAGAAUCCCUGUGAUCGUGGCCGAUGAUCUGACAGCCG CCAUCAACAAGGGCAUCCUCGUGACAGUGAACCCUAUCGCCUCCAC CAACGAUGACGAGGUGCUGAUCGAAGUGAACCCCCCCUUCGGCGAC UCCUACAUCAUCGUGGGCACAGGCGACAGCAGACUGACCUACCAGU GGCACAAAGAGGGCAGCAGCAUCGGCAAGCUGUUCACCCAGACCAU GAAGGGCGCCGAGAGACUGGCUGUGAUGGGAGAUGCCGCCUGGGAC UUUAGCAGCGCUGGCGGCUUCUUUACCAGCGUGGGCAAGGGAAUCC ACACCGUGUUCGGCAGCGCCUUCCAGGGACUGUUUGGCGGCCUGAG CUGGAUCACCAAAGUGAUCAUGGGCGCUGUGCUGAUCUGGGUGGGA AUCAACACCCGGAACAUGACCAUGAGCAUGUCCAUGAUCCUCGUGG GAGUGAUUAUGAUGUUCCUGAGCCUGGGCGUGGGCGCC YF_Sen2000_ AUGGAAACCCCUGCCCAGCUGCUGUUCCUGCUGCUGCUGUGGCUGC 90 prME_IgGksp CUGAUACCACCGGCGUGACACUCGUGCGGAAGAACAGAUGGCUGCU GCUGAACGUGACCAGCGAGGACCUGGGCAAGACCUUCUCUGUGGGC ACCGGCAACUGCACCACCAACAUCCUGGAAGCCAAGUACUGGUGCC CCGACAGCAUGGAGUACAACUGCCCCAACCUGAGCCCCAGAGAGGA ACCCGACGACAUCGACUGCUGGUGCUACGGCGUGGAAAACGUGCGG GUGGCCUACGGCAAGUGCGAUAGCGCCGGCAGAAGCAGAAGAAGCA GGCGGGCCAUCGACCUGCCCACCCACGAAAACCACGGCCUGAAAAC CCGGCAGGAAAAGUGGAUGACCGGCCGGAUGGGCGAGCGGCAGCUG CAGAAAAUCGAGAGAUGGCUCGUGCGCAACCCCUUCUUCGCCGUGA CCGCCCUGACAAUCGCCUACCUCGUGGGCAGCAACAUGACCCAGAG AGUCGUGAUCGCCCUGCUGGUGCUGGCUGUGGGCCCUGCCUAUAGC GCCCACUGUAUCGGCAUCACCGACCGGGACUUCAUCGAGGGCGUGC ACGGCGGAACAUGGGUGUCCGCUACCCUGGAACAGGAUAAGUGCGU GACCGUGAUGGCCCCCGACAAGCCCAGCCUGGACAUCAGCCUGGAA ACCGUGGCCAUUGACGGACCCGCCGAGGCCAGAAAAGUGUGCUACA ACGCCGUGCUGACCCACGUGAAGAUCAACGACAAGUGCCCCAGCAC CGGCGAAGCCCACCUGGCCGAAGAGAACGAGGGCGACAACGCCUGC AAGCGGACCUACAGCGAUAGAGGCUGGGGCAAUGGCUGCGGCCUGU UUGGCAAGGGCAGCAUCGUGGCCUGCGCCAAGUUCACCUGUGCCAA GAGCAUGAGCCUGUUCGAGGUGGACCAGACCAAGAUCCAGUACGUG AUCCGGGCUCAGCUGCACGUGGGCGCCAAGCAGGAAAACUGGAACA CCGACAUCAAGACCCUGAAGUUCGACGCCCUGAGCGGCUCCCAGGA AGCCGAGUUUACCGGCUAUGGCAAGGCCACCCUGGAAUGCCAGGUG CAGACCGCCGUGGACUUCGGCAACAGCUAUAUCGCCGAGAUGGAAA AAGAAAGCUGGAUCGUGGACCGGCAGUGGGCCCAGGAUCUGACACU GCCUUGGCAGUCUGGCUCUGGCGGAGUGUGGCGGGAAAUGCACCAC CUGGUGGAAUUCGAGCCUCCCCACGCCGCCACCAUUAGAGUGCUGG CCCUGGGCAAUCAGGAAGGCUCUCUGAAAACAGCCCUGACCGGCGC CAUGAGAGUGACCAAGGACACCAACGACAACAACCUGUACAAGCUG CACGGGGGGCACGUGUCCUGCAGAGUGAAACUGUCUGCCCUGACAC UGAAGGGCACCAGCUACAAGAUGUGCACCGACAAGAUGAGCUUCGU GAAGAACCCCACCGACACCGGCCACGGCACAGUCGUGAUGCAAGUG AAGGUGCCCAAGGGCGCUCCCUGCAGAAUCCCUGUGAUCGUGGCCG AUGAUCUGACAGCCGCCAUCAACAAGGGCAUCCUCGUGACAGUGAA CCCUAUCGCCUCCACCAACGAUGACGAGGUGCUGAUCGAAGUGAAC CCCCCCUUCGGCGACUCCUACAUCAUCGUGGGCACAGGCGACAGCA GACUGACCUACCAGUGGCACAAAGAGGGCAGCAGCAUCGGCAAGCU GUUCACCCAGACCAUGAAGGGCGCCGAGAGACUGGCUGUGAUGGGA GAUGCCGCCUGGGACUUUAGCAGCGCUGGCGGCUUCUUUACCAGCG UGGGCAAGGGAAUCCACACCGUGUUCGGCAGCGCCUUCCAGGGACU GUUUGGCGGCCUGAGCUGGAUCACCAAAGUGAUCAUGGGCGCUGUG CUGAUCUGGGUGGGAAUCAACACCCGGAACAUGACCAUGAGCAUGU CCAUGAUCCUCGUGGGAGUGAUUAUGAUGUUCCUGAGCCUGGGCGU GGGAGCC YF_17D_JEVsp_ AUGUGGCUGGUGUCCCUGGCCAUCGUGACAGCCUGUGCUGGCGCUG 91 SE_mutFL UGACACUCGUGCGGAAAAACAGAUGGCUGCUGCUGAACGUGACCAG CGAGGACCUGGGCAAGACCUUCUCUGUAGGCACCGGCAACUGCACC ACCAACAUCCUGGAAGCCAAGUACUGGUGCCCCGACAGCAUGGAGU ACAACUGCCCCAACCUGAGCCCCAGAGAGGAACCCGACGACAUCGA CUGCUGGUGCUACGGCGUGGAAAACGUGCGGGUGGCCUACGGCAAG UGCGAUAGCGCCGGCAGAAGCAGAAGAAGCAGGCGGGCCAUCGACC UGCCCACCCACGAAAACCACGGCCUGAAAACCCGGCAGGAAAAGUG GAUGACCGGCCGGAUGGGCGAGCGGCAGCUGCAGAAAAUUGAGCGG UGGUUUGUGCGGAACCCCUUCUUCGCCGUGACCGCCCUGACAAUCG CCUACCUCGUGGGCAGCAACAUGACCCAGAGAGUCGUGAUCGCCCU GCUGGUGCUGGCUGUGGGCCCUGCCUAUAGCGCCAUUGAUCUGCCU ACACACGAGAAUCAUGGGCUGAAAACAAGACAGGAAAAAUGGAUGA CUGGGCGCAUGGGAGAAAGACAGCUGCAGAAAAUCGAACGGUGGUU CGUGCGCAAUCCUUUUUUUGCUGUGACUGCUCUGACCAUUGCCUAU CUCGUGGGAUCCAAUAUGACACAGCGGGUCGUGAUUGCUCUGCUGG UGCUGGCAGUGGGACCCGCUUACUCCGCCCACUGUAUCGGCAUCAC CGACCGGGACUUCAUCGAGGGCGUGCACGGCGGAACAUGGGUGUCC GCUACCCUGGAACAGGAUAAGUGCGUGACCGUGAUGGCCCCCGACA AGCCCAGCCUGGACAUCAGCCUGGAAACCGUGGCCAUCGAUAGACC CGCCGAAGUGCGGAAAGUGUGCUACAACGCCGUGCUGACCCACGUG AAGAUCAACGACAAGUGCCCCAGAGAAGGCGAAGCCCACCUGGCCG AAGAGAACGAGGGCGACAACGCCUGCAAGCGGACCUACAGCGAUAG AGGCAGAGGCAAUGGCUGCGGCAGAUUUGGCAAGGGCAGCAUCGUG GCCUGCGCCAAGUUCACCUGUGCCAAGAGCAUGAGCCUGUUCGAGG UGGACCAGACCAAGAUCCAGUACGUGAUCCGGGCCCAGCUGCACGU GGGAGCCAAGCAGGAAAACUGGAACACCGACAUCAAGACCCUGAAG UUCGACGCCCUGAGCGGCUCCCAGGAAGUGGAAUUCAUCGGCUAUG GCAAGGCCACCCUGGAAUGUCAGGUGCAGACCGCCGUGGACUUCGG CAACAGCUAUAUCGCCGAGAUGGAAACCGAGAGCUGGAUCGUGGAC CGGCAGUGGGCUCAGGAUCUGACCCUGCCUUGGCAGUCUGGCUCUG GCGGAGUGUGGCGGGAAAUGCACCACCUGGUGGAAUUCGAGCCUCC CCACGCCGCCACCAUUAGAGUGCUGGCCCUGGGCAAUCAGGAAGGC UCUCUGAAAACAGCCCUGACCGGCGCCAUGAGAGUGACCAAGGACA CCAACGACAACAACCUGUACAAGCUGCAUGGCGGCCACGUGUCCUG CAGAGUGAAGCUGUCUGCCCUGACACUGAAGGGCACCAGCUACAAG AUCUGCACCGACAAGAUGUUCUUCGUGAAGAACCCCACCGACACCG GCCACGGCACAGUCGUGAUGCAAGUGAAGGUGUCCAAGGGCGCUCC CUGCCGGAUCCCUGUGAUCGUGGCCGAUGAUCUGACAGCCGCCAUC AACAAGGGCAUCCUCGUGACAGUGAACCCUAUCGCCUCCACCAACG AUGACGAGGUGCUGAUCGAAGUGAACCCCCCCUUCGGCGACUCCUA CAUCAUCGUGGGACGGGGCGACAGCAGACUGACCUACCAGUGGCAC AAAGAGGGCAGCAGCAUCGGCAAGCUGUUCACCCAGACCAUGAAGG GCGUGGAACGGCUGGCCGUGAUGGGAGAUACCGCCUGGGAUUUCUC UAGCGCUGGCGGCUUCUUCACCAGCGUGGGCAAGGGAAUCCACACC GUGUUCGGCAGCGCCUUCCAGGGACUGUUCGGCGGCCUGAACUGGA UCACCAAAGUGAUCAUGGGCGCUGUGCUGAUCUGGGUGGGAAUCAA CACCCGGAACAUGACCAUGAGCAUGUCCAUGAUCCUCGUGGGAGUG AUUAUGAUGUUCCUGAGCCUGGGUGUGGGCGCC YF_17D_IgGksp_ AUGGAAACCCCUGCCCAGCUGCUGUUCCUGCUGCUGCUGUGGCUGC 92 SE_mutFL CUGAUACCACCGGCGUGACACUCGUGCGGAAGAACAGAUGGCUGCU GCUGAACGUGACCAGCGAGGACCUGGGCAAGACCUUCUCUGUGGGC ACCGGCAACUGCACCACCAACAUCCUGGAAGCCAAGUACUGGUGCC CCGACAGCAUGGAGUACAACUGCCCCAACCUGAGCCCCAGAGAGGA ACCCGACGACAUCGACUGCUGGUGCUACGGCGUGGAAAACGUGCGG GUGGCCUACGGCAAGUGCGAUAGCGCCGGCAGAAGCAGAAGAAGCA GGCGGGCCAUCGACCUGCCCACCCACGAAAACCACGGCCUGAAAAC CCGGCAGGAAAAGUGGAUGACCGGCCGGAUGGGCGAGCGGCAGCUG CAGAAAAUUGAGCGGUGGUUUGUGCGGAACCCCUUCUUCGCCGUGA CCGCCCUGACAAUCGCCUACCUCGUGGGCAGCAACAUGACCCAGAG AGUCGUGAUCGCCCUGCUGGUGCUGGCUGUGGGCCCUGCCUAUAGC GCCAUUGAUCUGCCUACACACGAGAAUCAUGGGCUGAAAACAAGAC AGGAAAAAUGGAUGACUGGGCGCAUGGGAGAAAGACAGCUGCAGAA AAUCGAACGGUGGUUCGUGCGCAAUCCUUUUUUUGCUGUGACUGCU CUGACCAUUGCCUAUCUCGUGGGAUCCAAUAUGACACAGCGGGUCG UGAUUGCUCUGCUGGUGCUGGCAGUGGGACCCGCUUACUCCGCCCA CUGUAUCGGCAUCACCGACCGGGACUUCAUCGAGGGCGUGCACGGC GGAACAUGGGUGUCCGCUACCCUGGAACAGGAUAAGUGCGUGACCG UGAUGGCCCCCGACAAGCCCAGCCUGGACAUCAGCCUGGAAACCGU GGCCAUCGAUAGACCCGCCGAAGUGCGGAAAGUGUGCUACAACGCC GUGCUGACCCACGUGAAGAUCAACGACAAGUGCCCCAGAGAAGGCG AAGCCCACCUGGCCGAAGAGAACGAGGGCGACAACGCCUGCAAGCG GACCUACAGCGAUAGAGGCAGAGGCAAUGGCUGCGGCAGAUUUGGC AAGGGCAGCAUCGUGGCCUGCGCCAAGUUCACCUGUGCCAAGAGCA UGAGCCUGUUCGAGGUGGACCAGACCAAGAUCCAGUACGUGAUCCG GGCUCAGCUGCACGUGGGCGCCAAGCAGGAAAACUGGAACACCGAC AUCAAGACCCUGAAGUUCGACGCCCUGAGCGGCUCCCAGGAAGUGG AAUUCAUCGGCUAUGGCAAGGCCACCCUGGAAUGUCAGGUGCAGAC CGCCGUGGACUUCGGCAACAGCUAUAUCGCCGAGAUGGAAACCGAG AGCUGGAUCGUGGACCGGCAGUGGGCCCAGGAUCUGACACUGCCUU GGCAGUCUGGCUCUGGCGGAGUGUGGCGGGAAAUGCACCACCUGGU GGAAUUCGAGCCUCCCCACGCCGCCACCAUUAGAGUGCUGGCCCUG GGCAAUCAGGAAGGCUCUCUGAAAACAGCCCUGACCGGCGCCAUGA GAGUGACCAAGGACACCAACGACAACAACCUGUACAAGCUGCACGG GGGGCACGUGUCCUGCAGAGUGAAACUGUCUGCCCUGACACUGAAG GGCACCAGCUACAAGAUCUGCACCGACAAGAUGUUCUUCGUGAAGA ACCCCACCGACACCGGCCACGGCACAGUCGUGAUGCAAGUGAAGGU GUCCAAGGGCGCUCCCUGCCGGAUCCCUGUGAUCGUGGCCGAUGAU CUGACAGCCGCCAUCAACAAGGGCAUCCUCGUGACAGUGAACCCUA UCGCCUCCACCAACGAUGACGAGGUGCUGAUCGAAGUGAACCCCCC CUUCGGCGACUCCUACAUCAUCGUGGGACGGGGCGACAGCAGACUG ACCUACCAGUGGCACAAAGAGGGCAGCAGCAUCGGCAAGCUGUUCA CCCAGACCAUGAAGGGCGUGGAACGGCUGGCCGUGAUGGGAGAUAC CGCCUGGGAUUUCUCUAGCGCUGGCGGCUUCUUCACCAGCGUGGGC AAGGGAAUCCACACCGUGUUCGGCAGCGCCUUCCAGGGACUGUUCG GCGGCCUGAACUGGAUCACCAAAGUGAUCAUGGGCGCUGUGCUGAU CUGGGUGGGAAUCAACACCCGGAACAUGACCAUGAGCAUGUCCAUG AUCCUCGUGGGAGUGAUUAUGAUGUUCCUGAGCCUGGGCGUGGGUG CC YF_17D_JEVsp_ AUGUGGCUGGUGUCCCUGGCCAUCGUGACAGCCUGUGCUGGCGCUG 93 SE_no_polyNs_ UGACACUCGUGCGGAAGAACAGAUGGCUGCUGCUGAACGUGACCAG mutFL CGAGGACCUGGGCAAGACCUUCUCUGUAGGCACCGGCAACUGCACC ACCAACAUCCUGGAAGCCAAGUACUGGUGCCCCGACAGCAUGGAGU ACAACUGCCCCAACCUGAGCCCCAGAGAGGAACCCGACGACAUCGA CUGCUGGUGCUACGGCGUGGAAAACGUGCGGGUGGCCUACGGCAAG UGCGAUAGCGCCGGCAGAAGCAGAAGAAGCAGGCGGGCCAUCGACC UGCCCACCCACGAAAACCACGGCCUGAAAACCCGGCAGGAAAAGUG GAUGACCGGCCGGAUGGGCGAGCGGCAGCUGCAGAAAAUUGAGCGG UGGUUUGUGCGGAACCCCUUCUUCGCCGUGACCGCCCUGACAAUCG CCUACCUCGUGGGCAGCAACAUGACCCAGAGAGUCGUGAUCGCCCU GCUGGUGCUGGCUGUGGGCCCUGCCUAUAGCGCCAUUGAUCUGCCU ACACACGAGAAUCAUGGGCUGAAAACAAGACAGGAGAAAUGGAUGA CUGGGCGCAUGGGAGAAAGACAGCUGCAGAAAAUCGAACGGUGGUU CGUGCGCAAUCCUUUCUUUGCUGUGACUGCUCUGACCAUUGCCUAU CUCGUGGGAUCCAAUAUGACACAGCGGGUCGUGAUUGCUCUGCUGG UGCUGGCAGUGGGACCCGCUUACUCCGCCCACUGUAUCGGCAUCAC CGACCGGGACUUCAUCGAGGGCGUGCACGGCGGAACAUGGGUGUCC GCUACCCUGGAACAGGAUAAGUGCGUGACCGUGAUGGCCCCCGACA AGCCCAGCCUGGACAUCAGCCUGGAAACCGUGGCCAUCGAUAGACC CGCCGAAGUGCGGAAAGUGUGCUACAACGCCGUGCUGACCCACGUG AAGAUCAACGACAAGUGCCCCAGAGAAGGCGAAGCCCACCUGGCCG AAGAGAACGAGGGCGACAACGCCUGCAAGCGGACCUACAGCGAUAG AGGCAGAGGCAAUGGCUGCGGCAGAUUUGGCAAGGGCAGCAUCGUG GCCUGCGCCAAGUUCACCUGUGCCAAGAGCAUGAGCCUGUUCGAGG UGGACCAGACCAAGAUCCAGUACGUGAUCCGGGCCCAGCUGCACGU GGGAGCCAAGCAGGAAAACUGGAACACCGACAUCAAGACCCUGAAG UUCGACGCCCUGAGCGGCUCCCAGGAAGUGGAAUUCAUCGGCUAUG GCAAGGCCACCCUGGAAUGUCAGGUGCAGACCGCCGUGGACUUCGG CAACAGCUAUAUCGCCGAGAUGGAAACCGAGAGCUGGAUCGUGGAC CGGCAGUGGGCUCAGGAUCUGACCCUGCCUUGGCAGUCUGGCUCUG GCGGAGUGUGGCGGGAAAUGCACCACCUGGUGGAAUUCGAGCCUCC CCACGCCGCCACCAUUAGAGUGCUGGCCCUGGGCAAUCAGGAAGGC UCUCUGAAAACAGCCCUGACCGGCGCCAUGAGAGUGACCAAGGACA CCAACGACAACAACCUGUACAAGCUGCAUGGCGGCCACGUGUCCUG CAGAGUGAAGCUGUCUGCCCUGACACUGAAGGGCACCAGCUACAAG AUCUGCACCGACAAGAUGUUCUUCGUGAAGAACCCCACCGACACCG GCCACGGCACAGUCGUGAUGCAAGUGAAGGUGUCCAAGGGCGCUCC CUGCCGGAUCCCUGUGAUCGUGGCCGAUGAUCUGACAGCCGCCAUC AACAAGGGCAUCCUCGUGACAGUGAACCCUAUCGCCUCCACCAACG AUGACGAGGUGCUGAUCGAAGUGAAUCCUCCCUUCGGCGACUCCUA CAUCAUCGUGGGACGGGGCGACAGCAGACUGACCUACCAGUGGCAC AAAGAGGGCAGCAGCAUCGGCAAGCUGUUCACCCAGACCAUGAAGG GCGUGGAACGGCUGGCCGUGAUGGGAGAUACCGCCUGGGAUUUCUC UAGCGCUGGCGGCUUCUUCACCAGCGUGGGCAAGGGAAUCCACACC GUGUUCGGCAGCGCCUUCCAGGGACUGUUCGGCGGCCUGAACUGGA UCACCAAAGUGAUCAUGGGCGCUGUGCUGAUCUGGGUGGGAAUCAA CACCCGGAACAUGACCAUGAGCAUGUCCAUGAUCCUCGUGGGAGUG AUUAUGAUGUUCCUGAGCCUGGGUGUGGGCGCC YF_17D_JEVsp_ AUGUGGCUGGUGUCCCUGGCCAUCGUGACAGCCUGUGCUGGCGCUG 94 SE_no_polyNs UGACACUCGUGCGGAAGAACAGAUGGCUGCUGCUGAACGUGACCAG CGAGGACCUGGGCAAGACCUUCUCUGUAGGCACCGGCAACUGCACC ACCAACAUCCUGGAAGCCAAGUACUGGUGCCCCGACAGCAUGGAGU ACAACUGCCCCAACCUGAGCCCCAGAGAGGAACCCGACGACAUCGA CUGCUGGUGCUACGGCGUGGAAAACGUGCGGGUGGCCUACGGCAAG UGCGAUAGCGCCGGCAGAAGCAGAAGAAGCAGGCGGGCCAUCGACC UGCCCACCCACGAAAACCACGGCCUGAAAACCCGGCAGGAAAAGUG GAUGACCGGCCGGAUGGGCGAGCGGCAGCUGCAGAAAAUUGAGCGG UGGUUUGUGCGGAACCCCUUCUUCGCCGUGACCGCCCUGACAAUCG CCUACCUCGUGGGCAGCAACAUGACCCAGAGAGUCGUGAUCGCCCU GCUGGUGCUGGCUGUGGGCCCUGCCUAUAGCGCCAUUGAUCUGCCU ACACACGAGAAUCAUGGGCUGAAAACAAGACAGGAGAAAUGGAUGA CUGGGCGCAUGGGAGAAAGACAGCUGCAGAAAAUCGAACGGUGGUU CGUGCGCAAUCCUUUCUUUGCUGUGACUGCUCUGACCAUUGCCUAU CUCGUGGGAUCCAAUAUGACACAGCGGGUCGUGAUUGCUCUGCUGG UGCUGGCAGUGGGACCCGCUUACUCCGCCCACUGUAUCGGCAUCAC CGACCGGGACUUCAUCGAGGGCGUGCACGGCGGAACAUGGGUGUCC GCUACCCUGGAACAGGAUAAGUGCGUGACCGUGAUGGCCCCCGACA AGCCCAGCCUGGACAUCAGCCUGGAAACCGUGGCCAUCGAUAGACC CGCCGAAGUGCGGAAAGUGUGCUACAACGCCGUGCUGACCCACGUG AAGAUCAACGACAAGUGCCCCAGCACCGGCGAAGCCCACCUGGCCG AAGAGAACGAGGGCGACAACGCCUGCAAGCGGACCUACAGCGAUAG AGGCUGGGGCAAUGGCUGCGGCCUGUUUGGCAAGGGCAGCAUCGUG GCCUGCGCCAAGUUCACCUGUGCCAAGAGCAUGAGCCUGUUCGAGG UGGACCAGACCAAGAUCCAGUACGUGAUCCGGGCCCAGCUGCACGU GGGAGCCAAGCAGGAAAACUGGAACACCGACAUCAAGACCCUGAAG UUCGACGCCCUGAGCGGCUCCCAGGAAGUGGAAUUCAUCGGCUAUG GCAAGGCCACCCUGGAAUGUCAGGUGCAGACCGCCGUGGACUUCGG CAACAGCUAUAUCGCCGAGAUGGAAACCGAGAGCUGGAUCGUGGAC CGGCAGUGGGCUCAGGAUCUGACCCUGCCUUGGCAGUCUGGCUCUG GCGGAGUGUGGCGGGAAAUGCACCACCUGGUGGAAUUCGAGCCUCC CCACGCCGCCACCAUUAGAGUGCUGGCCCUGGGCAAUCAGGAAGGC UCUCUGAAAACAGCCCUGACCGGCGCCAUGAGAGUGACCAAGGACA CCAACGACAACAACCUGUACAAGCUGCAUGGCGGCCACGUGUCCUG CAGAGUGAAGCUGUCUGCCCUGACACUGAAGGGCACCAGCUACAAG AUCUGCACCGACAAGAUGUUCUUCGUGAAGAACCCCACCGACACCG GCCACGGCACAGUCGUGAUGCAAGUGAAGGUGUCCAAGGGCGCUCC CUGCCGGAUCCCUGUGAUCGUGGCCGAUGAUCUGACAGCCGCCAUC AACAAGGGCAUCCUCGUGACAGUGAACCCUAUCGCCUCCACCAACG AUGACGAGGUGCUGAUCGAAGUGAAUCCUCCCUUCGGCGACUCCUA CAUCAUCGUGGGACGGGGCGACAGCAGACUGACCUACCAGUGGCAC AAAGAGGGCAGCAGCAUCGGCAAGCUGUUCACCCAGACCAUGAAGG GCGUGGAACGGCUGGCCGUGAUGGGAGAUACCGCCUGGGAUUUCUC UAGCGCUGGCGGCUUCUUCACCAGCGUGGGCAAGGGAAUCCACACC GUGUUCGGCAGCGCCUUCCAGGGACUGUUCGGCGGCCUGAACUGGA UCACCAAAGUGAUCAUGGGCGCUGUGCUGAUCUGGGUGGGAAUCAA CACCCGGAACAUGACCAUGAGCAUGUCCAUGAUCCUCGUGGGAGUG AUUAUGAUGUUCCUGAGCCUGGGUGUGGGCGCC YF_17D_IgGksp_ AUGGAAACCCCUGCCCAGCUGCUGUUCCUGCUGCUGCUGUGGCUGC 95 SE_no_polyNs_ CUGAUACCACCGGCGUGACACUCGUGCGGAAGAACAGAUGGCUGCU mutFL GCUGAACGUGACCAGCGAGGACCUGGGCAAGACCUUCUCUGUGGGC ACCGGCAACUGCACCACCAACAUCCUGGAAGCCAAGUACUGGUGCC CCGACAGCAUGGAGUACAACUGCCCCAACCUGAGCCCCAGAGAGGA ACCCGACGACAUCGACUGCUGGUGCUACGGCGUGGAAAACGUGCGG GUGGCCUACGGCAAGUGCGAUAGCGCCGGCAGAAGCAGAAGAAGCA GGCGGGCCAUCGACCUGCCCACCCACGAAAACCACGGCCUGAAAAC CCGGCAGGAAAAGUGGAUGACCGGCCGGAUGGGCGAGCGGCAGCUG CAGAAAAUUGAGCGGUGGUUUGUGCGGAACCCCUUCUUCGCCGUGA CCGCCCUGACAAUCGCCUACCUCGUGGGCAGCAACAUGACCCAGAG AGUCGUGAUCGCCCUGCUGGUGCUGGCUGUGGGCCCUGCCUAUAGC GCCAUUGAUCUGCCUACACACGAGAAUCAUGGGCUGAAAACAAGAC AGGAGAAAUGGAUGACUGGGCGCAUGGGAGAAAGACAGCUGCAGAA AAUCGAACGGUGGUUCGUGCGCAAUCCGUUCUUUGCUGUGACUGCU CUGACCAUUGCCUAUCUCGUGGGAUCCAAUAUGACACAGCGGGUCG UGAUUGCUCUGCUGGUGCUGGCAGUGGGACCCGCUUACUCCGCCCA CUGUAUCGGCAUCACCGACCGGGACUUCAUCGAGGGCGUGCACGGC GGAACAUGGGUGUCCGCUACCCUGGAACAGGAUAAGUGCGUGACCG UGAUGGCCCCCGACAAGCCCAGCCUGGACAUCAGCCUGGAAACCGU GGCCAUCGAUAGACCCGCCGAAGUGCGGAAAGUGUGCUACAACGCC GUGCUGACCCACGUGAAGAUCAACGACAAGUGCCCCAGAGAAGGCG AAGCCCACCUGGCCGAAGAGAACGAGGGCGACAACGCCUGCAAGCG GACCUACAGCGAUAGAGGCAGAGGCAAUGGCUGCGGCAGAUUUGGC AAGGGCAGCAUCGUGGCCUGCGCCAAGUUCACCUGUGCCAAGAGCA UGAGCCUGUUCGAGGUGGACCAGACCAAGAUCCAGUACGUGAUCCG GGCUCAGCUGCACGUGGGCGCCAAGCAGGAAAACUGGAACACCGAC AUCAAGACCCUGAAGUUCGACGCCCUGAGCGGCUCCCAGGAAGUGG AAUUCAUCGGCUAUGGCAAGGCCACCCUGGAAUGUCAGGUGCAGAC CGCCGUGGACUUCGGCAACAGCUAUAUCGCCGAGAUGGAAACCGAG AGCUGGAUCGUGGACCGGCAGUGGGCCCAGGAUCUGACACUGCCUU GGCAGUCUGGCUCUGGCGGAGUGUGGCGGGAAAUGCACCACCUGGU GGAAUUCGAGCCUCCCCACGCCGCCACCAUUAGAGUGCUGGCCCUG GGCAAUCAGGAAGGCUCUCUGAAAACAGCCCUGACCGGCGCCAUGA GAGUGACCAAGGACACCAACGACAACAACCUGUACAAGCUGCACGG GGGGCACGUGUCCUGCAGAGUGAAACUGUCUGCCCUGACACUGAAG GGCACCAGCUACAAGAUCUGCACCGACAAGAUGUUCUUCGUGAAGA ACCCCACCGACACCGGCCACGGCACAGUCGUGAUGCAAGUGAAGGU GUCCAAGGGCGCUCCCUGCCGGAUCCCUGUGAUCGUGGCCGAUGAU CUGACAGCCGCCAUCAACAAGGGCAUCCUCGUGACAGUGAACCCUA UCGCCUCCACCAACGAUGACGAGGUGCUGAUCGAAGUGAACCCUCC UUUCGGCGACUCCUACAUCAUCGUGGGACGGGGCGACAGCAGACUG ACCUACCAGUGGCACAAAGAGGGCAGCAGCAUCGGCAAGCUGUUCA CCCAGACCAUGAAGGGCGUGGAACGGCUGGCCGUGAUGGGAGAUAC CGCCUGGGAUUUCUCUAGCGCUGGCGGCUUCUUCACCAGCGUGGGC AAGGGAAUCCACACCGUGUUCGGCAGCGCCUUCCAGGGACUGUUCG GCGGCCUGAACUGGAUCACCAAAGUGAUCAUGGGCGCUGUGCUGAU CUGGGUGGGAAUCAACACCCGGAACAUGACCAUGAGCAUGUCCAUG AUCCUCGUGGGAGUGAUUAUGAUGUUCCUGAGCCUGGGCGUGGGUG CC YF_17D_IgGksp_ AUGGAAACCCCUGCCCAGCUGCUGUUCCUGCUGCUGCUGUGGCUGC 96 SE_no_polyNs CUGAUACCACCGGCGUGACACUCGUGCGGAAGAACAGAUGGCUGCU GCUGAACGUGACCAGCGAGGACCUGGGCAAGACCUUCUCUGUGGGC ACCGGCAACUGCACCACCAACAUCCUGGAAGCCAAGUACUGGUGCC CCGACAGCAUGGAGUACAACUGCCCCAACCUGAGCCCCAGAGAGGA ACCCGACGACAUCGACUGCUGGUGCUACGGCGUGGAAAACGUGCGG GUGGCCUACGGCAAGUGCGAUAGCGCCGGCAGAAGCAGAAGAAGCA GGCGGGCCAUCGACCUGCCCACCCACGAAAACCACGGCCUGAAAAC CCGGCAGGAAAAGUGGAUGACCGGCCGGAUGGGCGAGCGGCAGCUG CAGAAAAUUGAGCGGUGGUUUGUGCGGAACCCCUUCUUCGCCGUGA CCGCCCUGACAAUCGCCUACCUCGUGGGCAGCAACAUGACCCAGAG AGUCGUGAUCGCCCUGCUGGUGCUGGCUGUGGGCCCUGCCUAUAGC GCCAUUGAUCUGCCUACACACGAGAAUCAUGGGCUGAAAACAAGAC AGGAGAAAUGGAUGACUGGGCGCAUGGGAGAAAGACAGCUGCAGAA AAUCGAACGGUGGUUCGUGCGCAAUCCGUUCUUUGCUGUGACUGCU CUGACCAUUGCCUAUCUCGUGGGAUCCAAUAUGACACAGCGGGUCG UGAUUGCUCUGCUGGUGCUGGCAGUGGGACCCGCUUACUCCGCCCA CUGUAUCGGCAUCACCGACCGGGACUUCAUCGAGGGCGUGCACGGC GGAACAUGGGUGUCCGCUACCCUGGAACAGGAUAAGUGCGUGACCG UGAUGGCCCCCGACAAGCCCAGCCUGGACAUCAGCCUGGAAACCGU GGCCAUCGAUAGACCCGCCGAAGUGCGGAAAGUGUGCUACAACGCC GUGCUGACCCACGUGAAGAUCAACGACAAGUGCCCCAGCACCGGCG AAGCCCACCUGGCCGAAGAGAACGAGGGCGACAACGCCUGCAAGCG GACCUACAGCGAUAGAGGCUGGGGCAAUGGCUGCGGCCUGUUUGGC AAGGGCAGCAUCGUGGCCUGCGCCAAGUUCACCUGUGCCAAGAGCA UGAGCCUGUUCGAGGUGGACCAGACCAAGAUCCAGUACGUGAUCCG GGCUCAGCUGCACGUGGGCGCCAAGCAGGAAAACUGGAACACCGAC AUCAAGACCCUGAAGUUCGACGCCCUGAGCGGCUCCCAGGAAGUGG AAUUCAUCGGCUAUGGCAAGGCCACCCUGGAAUGUCAGGUGCAGAC CGCCGUGGACUUCGGCAACAGCUAUAUCGCCGAGAUGGAAACCGAG AGCUGGAUCGUGGACCGGCAGUGGGCCCAGGAUCUGACACUGCCUU GGCAGUCUGGCUCUGGCGGAGUGUGGCGGGAAAUGCACCACCUGGU GGAAUUCGAGCCUCCCCACGCCGCCACCAUUAGAGUGCUGGCCCUG GGCAAUCAGGAAGGCUCUCUGAAAACAGCCCUGACCGGCGCCAUGA GAGUGACCAAGGACACCAACGACAACAACCUGUACAAGCUGCACGG GGGGCACGUGUCCUGCAGAGUGAAACUGUCUGCCCUGACACUGAAG GGCACCAGCUACAAGAUCUGCACCGACAAGAUGUUCUUCGUGAAGA ACCCCACCGACACCGGCCACGGCACAGUCGUGAUGCAAGUGAAGGU GUCCAAGGGCGCUCCCUGCCGGAUCCCUGUGAUCGUGGCCGAUGAU CUGACAGCCGCCAUCAACAAGGGCAUCCUCGUGACAGUGAACCCUA UCGCCUCCACCAACGAUGACGAGGUGCUGAUCGAAGUGAACCCUCC UUUCGGCGACUCCUACAUCAUCGUGGGACGGGGCGACAGCAGACUG ACCUACCAGUGGCACAAAGAGGGCAGCAGCAUCGGCAAGCUGUUCA CCCAGACCAUGAAGGGCGUGGAACGGCUGGCCGUGAUGGGAGAUAC CGCCUGGGAUUUCUCUAGCGCUGGCGGCUUCUUCACCAGCGUGGGC AAGGGAAUCCACACCGUGUUCGGCAGCGCCUUCCAGGGACUGUUCG GCGGCCUGAACUGGAUCACCAAAGUGAUCAUGGGCGCUGUGCUGAU CUGGGUGGGAAUCAACACCCGGAACAUGACCAUGAGCAUGUCCAUG AUCCUCGUGGGAGUGAUUAUGAUGUUCCUGAGCCUGGGCGUGGGUG CC

TABLE 22 YFV Amino Acid Sequences SEQ ID Description Sequence NO: YF_Con_prME_ METPAQLLFLLLLWLPDTTGVTLVRKNRWLLLNVTSEDLGKTFSVGT  97 Sen2000_IgGksp GNCTTNILEAKYWCPDSMEYNCPNLSPREEPDDIDCWCYGVENVRVA (signal peptide YGKCDSAGRSRRSRRAIDLPTHENHGLKTRQEKWMTGRMGERQLQKI underlined: SEQ ID ERWLVRNPFFAVTALTIAYLVGSNMTQRVVIALLVLAVGPAYSAHCIG NO: 423) ITDRDFIEGVHGGTWVSATLEQDKCVTVMAPDKPSLDISLETVAIDGPA EARKVCYNAVLTHVKINDKCPSTGEAHLAEENEGDNACKRTYSDRG WGNGCGLFGKGSIVACAKFTCAKSMSLFEVDQTKIQYVIRAQLHVGA KQENWNTDIKTLKFDALSGSQEAEFTGYGKATLECQVQTAVDFGNSYI AEMEKESWIVDRQWAQDLTLPWQSGSGGVWREMHHLVEFEPPHAAT IRVLALGNQEGSLKTALTGAMRVTKDTNDNNLYKLHGGHVSCRVKLS ALTLKGTSYKMCTDKMSFVKNPTDTGHGTVVMQVKVPKGAPCRIPVI VADDLTAAINKGILVTVNPIASTNDDEVLIEVNPPFGDSYIIVGTGDSRL TYQWHKEGSSIGKLFTQTMKGAERLAVMGDAAWDFSSAGGFFTSVG KGIHTVFGSAFQGLFGGLSWITKVIMGAVLIWVGINTRNMTMSMSMIL VGVIMMFLSLGVGA YF_Con_prME_ MWLVSLAIVTACAGAVTLVRKNRWLLLNVTSEDLGKTFSVGTGNCTT  98 Sen2000_JEVsp NILEAKYWCPDSMEYNCPNLSPREEPDDIDCWCYGVENVRVAYGKCD (signal peptide SAGRSRRSRRAIDLPTHENHGLKTRQEKWMTGRMGERQLQKIERWLV underlined: SEQ ID RNPFFAVTALTIAYLVGSNMTQRVVIALLVLAVGPAYSAHCIGITDRDF NO: 15) IEGVHGGTWVSATLEQDKCVTVMAPDKPSLDISLETVAIDGPAEARKV CYNAVLTHVKINDKCPSTGEAHLAEENEGDNACKRTYSDRGWGNGC GLFGKGSIVACAKFTCAKSMSLFEVDQTKIQYVIRAQLHVGAKQENW NTDIKTLKFDALSGSQEAEFTGYGKATLECQVQTAVDFGNSYIAEMEK ESWIVDRQWAQDLTLPWQSGSGGVWREMHHLVEFEPPHAATIRVLAL GNQEGSLKTALTGAMRVTKDTNDNNLYKLHGGHVSCRVKLSALTLK GTSYKMCTDKMSFVKNPTDTGHGTVVMQVKVPKGAPCRIPVIVADDL TAAINKGILVTVNPIASTNDDEVLIEVNPPFGDSYIIVGTGDSRLTYQWH KEGSSIGKLFTQTMKGAERLAVMGDAAWDFSSAGGFFTSVGKGIHTV FGSAFQGLFGGLSWITKVIMGAVLIWVGINTRNMTMSMSMILVGVIM MFLSLGVGA YF_Con_prME_ MWLVSLAIVTACAGAVTLVRKNRWLLLNVTSEDLGKTFSVGTGNCTT  99 Sen2000_JEVsp_ NILEAKYWCPDSMEYNCPNLSPREEPDDIDCWCYGVENVRVAYGKCD N153T (signal peptide SAGRSRRSRRAIDLPTHENHGLKTRQEKWMTGRMGERQLQKIERWLV underlined: SEQ ID RNPFFAVTALTIAYLVGSNMTQRVVIALLVLAVGPAYSAHCIGITDRDF NO: 15) IEGVHGGTWVSATLEQDKCVTVMAPDKPSLDISLETVAIDGPAEARKV CYNAVLTHVKINDKCPSTGEAHLAEENEGDNACKRTYSDRGWGNGC GLFGKGSIVACAKFTCAKSMSLFEVDQTKIQYVIRAQLHVGAKQENW TTDIKTLKFDALSGSQEAEFTGYGKATLECQVQTAVDFGNSYIAEMEK ESWIVDRQWAQDLTLPWQSGSGGVWREMHHLVEFEPPHAATIRVLAL GNQEGSLKTALTGAMRVTKDTNDNNLYKLHGGHVSCRVKLSALTLK GTSYKMCTDKMSFVKNPTDTGHGTVVMQVKVPKGAPCRIPVIVADDL TAAINKGILVTVNPIASTNDDEVLIEVNPPFGDSYIIVGTGDSRLTYQWH KEGSSIGKLFTQTMKGAERLAVMGDAAWDFSSAGGFFTSVGKGIHTV FGSAFQGLFGGLSWITKVIMGAVLIWVGINTRNMTMSMSMILVGVIM MFLSLGVGA 17D_vaccine_ MWLVSLAIVTACAGAVTLVRKNRWLLLNVTSEDLGKTFSVGTGNCTT 100 prME_JEVsp (signal NILEAKYWCPDSMEYNCPNLSPREEPDDIDCWCYGVENVRVAYGKCD peptide underlined: SAGRSRRSRRAIDLPTHENHGLKTRQEKWMTGRMGERQLQKIERWFV SEQ ID NO: 15) RNPFFAVTALTIAYLVGSNMTQRVVIALLVLAVGPAYSAIDLPTHENH GLKTRQEKWMTGRMGERQLQKIERWFVRNPFFAVTALTIAYLVGSN MTQRVVIALLVLAVGPAYSAHCIGITDRDFIEGVHGGTWVSATLEQDK CVTVMAPDKPSLDISLETVAIDRPAEVRKVCYNAVLTHVKINDKCPST GEAHLAEENEGDNACKRTYSDRGWGNGCGLFGKGSIVACAKFTCAKS MSLFEVDQTKIQYVIRAQLHVGAKQENWNTDIKTLKFDALSGSQEVEF IGYGKATLECQVQTAVDFGNSYIAEMETESWIVDRQWAQDLTLPWQS GSGGVWREMHHLVEFEPPHAATIRVLALGNQEGSLKTALTGAMRVTK DTNDNNLYKLHGGHVSCRVKLSALTLKGTSYKICTDKMFFVKNPTDT GHGTVVMQVKVSKGAPCRIPVIVADDLTAAINKGILVTVNPIASTNDD EVLIEVNPPFGDSYIIVGRGDSRLTYQWHKEGSSIGKLFTQTMKGVERL AVMGDTAWDFSSAGGFFTSVGKGIHTVFGSAFQGLFGGLNWITKVIM GAVLIWVGINTRNMTMSMSMILVGVIMMFLSLGVGA 17D_vaccine_ METPAQLLFLLLLWLPDTTGVTLVRKNRWLLLNVTSEDLGKTFSVGT 101 prME_IgGKsp (signal GNCTTNILEAKYWCPDSMEYNCPNLSPREEPDDIDCWCYGVENVRVA peptide underlined: YGKCDSAGRSRRSRRAIDLPTHENHGLKTRQEKWMTGRMGERQLQKI SEQ ID NO: 423) ERWFVRNPFFAVTALTIAYLVGSNMTQRVVIALLVLAVGPAYSAIDLP THENHGLKTRQEKWMTGRMGERQLQKIERWFVRNPFFAVTALTIAYL VGSNMTQRVVIALLVLAVGPAYSAHCIGITDRDFIEGVHGGTWVSATL EQDKCVTVMAPDKPSLDISLETVAIDRPAEVRKVCYNAVLTHVKINDK CPSTGEAHLAEENEGDNACKRTYSDRGWGNGCGLFGKGSIVACAKFT CAKSMSLFEVDQTKIQYVIRAQLHVGAKQENWNTDIKTLKFDALSGS QEVEFIGYGKATLECQVQTAVDFGNSYIAEMETESWIVDRQWAQDLT LPWQSGSGGVWREMHHLVEFEPPHAATIRVLALGNQEGSLKTALTGA MRVTKDTNDNNLYKLHGGHVSCRVKLSALTLKGTSYKICTDKMFFVK NPTDTGHGTVVMQVKVSKGAPCRIPVIVADDLTAAINKGILVTVNPIAS TNDDEVLIEVNPPFGDSYIIVGRGDSRLTYQWHKEGSSIGKLFTQTMKG VERLAVMGDTAWDFSSAGGFFTSVGKGIHTVFGSAFQGLFGGLNWIT KVIMGAVLIWVGINTRNMTMSMSMILVGVIMMFLSLGVGA YF_Con_prME_ VTLVRKNRWLLLNVTSEDLGKTFSVGTGNCTTNILEAKYWCPDSMEY 102 Sen2000_IgGksp (no NCPNLSPREEPDDIDCWCYGVENVRVAYGKCDSAGRSRRSRRAIDLPT signal peptide) HENHGLKTRQEKWMTGRMGERQLQKIERWLVRNPFFAVTALTIAYLV GSNMTQRVVIALLVLAVGPAYSAHCIGITDRDFIEGVHGGTWVSATLE QDKCVTVMAPDKPSLDISLETVAIDGPAEARKVCYNAVLTHVKINDKC PSTGEAHLAEENEGDNACKRTYSDRGWGNGCGLFGKGSIVACAKFTC AKSMSLFEVDQTKIQYVIRAQLHVGAKQENWNTDIKTLKFDALSGSQE AEFTGYGKATLECQVQTAVDFGNSYIAEMEKESWIVDRQWAQDLTLP WQSGSGGVWREMHHLVEFEPPHAATIRVLALGNQEGSLKTALTGAM RVTKDTNDNNLYKLHGGHVSCRVKLSALTLKGTSYKMCTDKMSFVK NPTDTGHGTVVMQVKVPKGAPCRIPVIVADDLTAAINKGILVTVNPIAS TNDDEVLIEVNPPFGDSYIIVGTGDSRLTYQWHKEGSSIGKLFTQTMKG AERLAVMGDAAWDFSSAGGFFTSVGKGIHTVFGSAFQGLFGGLSWIT KVIMGAVLIWVGINTRNMTMSMSMILVGVIMMFLSLGVGA YF_Con_prME_ VTLVRKNRWLLLNVTSEDLGKTFSVGTGNCTTNILEAKYWCPDSMEY 103 Sen2000_JEVsp (no NCPNLSPREEPDDIDCWCYGVENVRVAYGKCDSAGRSRRSRRAIDLPT signal peptide) HENHGLKTRQEKWMTGRMGERQLQKIERWLVRNPFFAVTALTIAYLV GSNMTQRVVIALLVLAVGPAYSAHCIGITDRDFIEGVHGGTWVSATLE QDKCVTVMAPDKPSLDISLETVAIDGPAEARKVCYNAVLTHVKINDKC PSTGEAHLAEENEGDNACKRTYSDRGWGNGCGLFGKGSIVACAKFTC AKSMSLFEVDQTKIQYVIRAQLHVGAKQENWNTDIKTLKFDALSGSQE AEFTGYGKATLECQVQTAVDFGNSYIAEMEKESWIVDRQWAQDLTLP WQSGSGGVWREMHHLVEFEPPHAATIRVLALGNQEGSLKTALTGAM RVTKDTNDNNLYKLHGGHVSCRVKLSALTLKGTSYKMCTDKMSFVK NPTDTGHGTVVMQVKVPKGAPCRIPVIVADDLTAAINKGILVTVNPIAS TNDDEVLIEVNPPFGDSYIIVGTGDSRLTYQWHKEGSSIGKLFTQTMKG AERLAVMGDAAWDFSSAGGFFTSVGKGIHTVFGSAFQGLFGGLSWIT KVIMGAVLIWVGINTRNMTMSMSMILVGVIMMFLSLGVGA YF_Con_prME_ VTLVRKNRWLLLNVTSEDLGKTFSVGTGNCTTNILEAKYWCPDSMEY 104 Sen2000_JEVsp_ NCPNLSPREEPDDIDCWCYGVENVRVAYGKCDSAGRSRRSRRAIDLPT N153T (no signal HENHGLKTRQEKWMTGRMGERQLQKIERWLVRNPFFAVTALTIAYLV peptide) GSNMTQRVVIALLVLAVGPAYSAHCIGITDRDFIEGVHGGTWVSATLE QDKCVTVMAPDKPSLDISLETVAIDGPAEARKVCYNAVLTHVKINDKC PSTGEAHLAEENEGDNACKRTYSDRGWGNGCGLFGKGSIVACAKFTC AKSMSLFEVDQTKIQYVIRAQLHVGAKQENWTTDIKTLKFDALSGSQE AEFTGYGKATLECQVQTAVDFGNSYIAEMEKESWIVDRQWAQDLTLP WQSGSGGVWREMHHLVEFEPPHAATIRVLALGNQEGSLKTALTGAM RVTKDTNDNNLYKLHGGHVSCRVKLSALTLKGTSYKMCTDKMSFVK NPTDTGHGTVVMQVKVPKGAPCRIPVIVADDLTAAINKGILVTVNPIAS TNDDEVLIEVNPPFGDSYIIVGTGDSRLTYQWHKEGSSIGKLFTQTMKG AERLAVMGDAAWDFSSAGGFFTSVGKGIHTVFGSAFQGLFGGLSWIT KVIMGAVLIWVGINTRNMTMSMSMILVGVIMMFLSLGVGA 17D_vaccine_ VTLVRKNRWLLLNVTSEDLGKTFSVGTGNCTTNILEAKYWCPDSMEY 105 prME_JEVsp (no signal NCPNLSPREEPDDIDCWCYGVENVRVAYGKCDSAGRSRRSRRAIDLPT peptide) HENHGLKTRQEKWMTGRMGERQLQKIERWFVRNPFFAVTALTIAYLV GSNMTQRVVIALLVLAVGPAYSAIDLPTHENHGLKTRQEKWMTGRM GERQLQKIERWFVRNPFFAVTALTIAYLVGSNMTQRVVIALLVLAVGP AYSAHCIGITDRDFIEGVHGGTWVSATLEQDKCVTVMAPDKPSLDISL ETVAIDRPAEVRKVCYNAVLTHVKINDKCPSTGEAHLAEENEGDNAC KRTYSDRGWGNGCGLFGKGSIVACAKFTCAKSMSLFEVDQTKIQYVIR AQLHVGAKQENWNTDIKTLKFDALSGSQEVEFIGYGKATLECQVQTA VDFGNSYIAEMETESWIVDRQWAQDLTLPWQSGSGGVWREMHHLVE PEPPHAATIRVLALGNQEGSLKTALTGAMRVTKDTNDNNLYKLHGGH VSCRVKLSALTLKGTSYKICTDKMFFVKNPTDTGHGTVVMQVKVSKG APCRIPVIVADDLTAAINKGILVTVNPIASTNDDEVLIEVNPPFGDSYIIV GRGDSRLTYQWHKEGSSIGKLFTQTMKGVERLAVMGDTAWDFSSAG GFFTSVGKGIHTVFGSAFQGLFGGLNWITKVIMGAVLIWVGINTRNMT MSMSMILVGVIMMFLSLGVGA 17D_vaccine_ VTLVRKNRWLLLNVTSEDLGKTFSVGTGNCTTNILEAKYWCPDSMEY 106 prME_IgGKsp (no NCPNLSPREEPDDIDCWCYGVENVRVAYGKCDSAGRSRRSRRAIDLPT signal peptide) HENHGLKTRQEKWMTGRMGERQLQKIERWFVRNPFFAVTALTIAYLV GSNMTQRVVIALLVLAVGPAYSAIDLPTHENHGLKTRQEKWMTGRM GERQLQKIERWFVRNPFFAVTALTIAYLVGSNMTQRVVIALLVLAVGP AYSAHCIGITDRDFIEGVHGGTWVSATLEQDKCVTVMAPDKPSLDISL ETVAIDRPAEVRKVCYNAVLTHVKINDKCPSTGEAHLAEENEGDNAC KRTYSDRGWGNGCGLFGKGSIVACAKFTCAKSMSLFEVDQTKIQYVIR AQLHVGAKQENWNTDIKTLKFDALSGSQEVEFIGYGKATLECQVQTA VDFGNSYIAEMETESWIVDRQWAQDLTLPWQSGSGGVWREMHHLVE PEPPHAATIRVLALGNQEGSLKTALTGAMRVTKDTNDNNLYKLHGGH VSCRVKLSALTLKGTSYKICTDKMFFVKNPTDTGHGTVVMQVKVSKG APCRIPVIVADDLTAAINKGILVTVNPIASTNDDEVLIEVNPPFGDSYIIV GRGDSRLTYQWHKEGSSIGKLFTQTMKGVERLAVMGDTAWDFSSAG GFFTSVGKGIHTVFGSAFQGLFGGLNWITKVIMGAVLIWVGINTRNMT MSMSMILVGVIMMFLSLGVGA YF_Sen2000_ MWLVSLAIVTACAGAVTLVRKNRWLLLNVTSEDLGKTFSVGTGNCTT 107 JEVsp_N153T NILEAKYWCPDSMEYNCPNLSPREEPDDIDCWCYGVENVRVAYGKCD SAGRSRRSRRAIDLPTHENHGLKTRQEKWMTGRMGERQLQKIERWLV RNPFFAVTALTIAYLVGSNMTQRVVIALLVLAVGPAYSAHCIGITDRDF IEGVHGGTWVSATLEQDKCVTVMAPDKPSLDISLETVAIDGPAEARKV CYNAVLTHVKINDKCPSTGEAHLAEENEGDNACKRTYSDRGWGNGC GLFGKGSIVACAKFTCAKSMSLFEVDQTKIQYVIRAQLHVGAKQENW TTDIKTLKFDALSGSQEAEFTGYGKATLECQVQTAVDFGNSYIAEMEK ESWIVDRQWAQDLTLPWQSGSGGVWREMHHLVEFEPPHAATIRVLAL GNQEGSLKTALTGAMRVTKDTNDNNLYKLHGGHVSCRVKLSALTLK GTSYKMCTDKMSFVKNPTDTGHGTVVMQVKVPKGAPCRIPVIVADDL TAAINKGILVTVNPIASTNDDEVLIEVNPPFGDSYIIVGTGDSRLTYQWH KEGSSIGKLFTQTMKGAERLAVMGDAAWDFSSAGGFFTSVGKGIHTV FGSAFQGLFGGLSWITKVIMGAVLIWVGINTRNMTMSMSMILVGVIM MFLSLGVGA YF_17D_IgGksp_ METPAQLLFLLLLWLPDTTGVTLVRKNRWLLLNVTSEDLGKTFSVGT 108 SE GNCTTNILEAKYWCPDSMEYNCPNLSPREEPDDIDCWCYGVENVRVA YGKCDSAGRSRRSRRAIDLPTHENHGLKTRQEKWMTGRMGERQLQKI ERWFVRNPFFAVTALTIAYLVGSNMTQRVVIALLVLAVGPAYSAIDLP THENHGLKTRQEKWMTGRMGERQLQKIERWFVRNPFFAVTALTIAYL VGSNMTQRVVIALLVLAVGPAYSAHCIGITDRDFIEGVHGGTWVSATL EQDKCVTVMAPDKPSLDISLETVAIDRPAEVRKVCYNAVLTHVKINDK CPSTGEAHLAEENEGDNACKRTYSDRGWGNGCGLFGKGSIVACAKFT CAKSMSLFEVDQTKIQYVIRAQLHVGAKQENWNTDIKTLKFDALSGS QEVEFIGYGKATLECQVQTAVDFGNSYIAEMETESWIVDRQWAQDLT LPWQSGSGGVWREMHHLVEFEPPHAATIRVLALGNQEGSLKTALTGA MRVTKDTNDNNLYKLHGGHVSCRVKLSALTLKGTSYKICTDKMFFVK NPTDTGHGTVVMQVKVSKGAPCRIPVIVADDLTAAINKGILVTVNPIAS TNDDEVLIEVNPPFGDSYIIVGRGDSRLTYQWHKEGSSIGKLFTQTMKG VERLAVMGDTAWDFSSAGGFFTSVGKGIHTVFGSAFQGLFGGLNWIT KVIMGAVLIWVGINTRNMTMSMSMILVGVIMMFLSLGVGA YF_17D_JEVsp_ MWLVSLAIVTACAGAVTLVRKNRWLLLNVTSEDLGKTFSVGTGNCTT 109 SE NILEAKYWCPDSMEYNCPNLSPREEPDDIDCWCYGVENVRVAYGKCD SAGRSRRSRRAIDLPTHENHGLKTRQEKWMTGRMGERQLQKIERWFV RNPFFAVTALTIAYLVGSNMTQRVVIALLVLAVGPAYSAIDLPTHENH GLKTRQEKWMTGRMGERQLQKIERWFVRNPFFAVTALTIAYLVGSN MTQRVVIALLVLAVGPAYSAHCIGITDRDFIEGVHGGTWVSATLEQDK CVTVMAPDKPSLDISLETVAIDRPAEVRKVCYNAVLTHVKINDKCPST GEAHLAEENEGDNACKRTYSDRGWGNGCGLFGKGSIVACAKFTCAKS MSLFEVDQTKIQYVIRAQLHVGAKQENWNTDIKTLKFDALSGSQEVEF IGYGKATLECQVQTAVDFGNSYIAEMETESWIVDRQWAQDLTLPWQS GSGGVWREMHHLVEFEPPHAATIRVLALGNQEGSLKTALTGAMRVTK DTNDNNLYKLHGGHVSCRVKLSALTLKGTSYKICTDKMFFVKNPTDT GHGTVVMQVKVSKGAPCRIPVIVADDLTAAINKGILVTVNPIASTNDD EVLIEVNPPFGDSYIIVGRGDSRLTYQWHKEGSSIGKLFTQTMKGVERL AVMGDTAWDFSSAGGFFTSVGKGIHTVFGSAFQGLFGGLNWITKVIM GAVLIWVGINTRNMTMSMSMILVGVIMMFLSLGVGA YF_SEN2000_ MWLVSLAIVTACAGAVTLVRKNRWLLLNVTSEDLGKTFSVGTGNCTT 110 prME_JEVsp NILEAKYWCPDSMEYNCPNLSPREEPDDIDCWCYGVENVRVAYGKCD SAGRSRRSRRAIDLPTHENHGLKTRQEKWMTGRMGERQLQKIERWLV RNPFFAVTALTIAYLVGSNMTQRVVIALLVLAVGPAYSAHCIGITDRDF IEGVHGGTWVSATLEQDKCVTVMAPDKPSLDISLETVAIDGPAEARKV CYNAVLTHVKINDKCPSTGEAHLAEENEGDNACKRTYSDRGWGNGC GLFGKGSIVACAKFTCAKSMSLFEVDQTKIQYVIRAQLHVGAKQENW NTDIKTLKFDALSGSQEAEFTGYGKATLECQVQTAVDFGNSYIAEMEK ESWIVDRQWAQDLTLPWQSGSGGVWREMHHLVEFEPPHAATIRVLAL GNQEGSLKTALTGAMRVTKDTNDNNLYKLHGGHVSCRVKLSALTLK GTSYKMCTDKMSFVKNPTDTGHGTVVMQVKVPKGAPCRIPVIVADDL TAAINKGILVTVNPIASTNDDEVLIEVNPPFGDSYIIVGTGDSRLTYQWH KEGSSIGKLFTQTMKGAERLAVMGDAAWDFSSAGGFFTSVGKGIHTV FGSAFQGLFGGLSWITKVIMGAVLIWVGINTRNMTMSMSMILVGVIM MFLSLGVGA YF_Sen2000_ METPAQLLFLLLLWLPDTTGVTLVRKNRWLLLNVTSEDLGKTFSVGT 111 prME_IgGksp GNCTTNILEAKYWCPDSMEYNCPNLSPREEPDDIDCWCYGVENVRVA YGKCDSAGRSRRSRRAIDLPTHENHGLKTRQEKWMTGRMGERQLQKI ERWLVRNPFFAVTALTIAYLVGSNMTQRVVIALLVLAVGPAYSAHCIG ITDRDFIEGVHGGTWVSATLEQDKCVTVMAPDKPSLDISLETVAIDGPA EARKVCYNAVLTHVKINDKCPSTGEAHLAEENEGDNACKRTYSDRG WGNGCGLFGKGSIVACAKFTCAKSMSLFEVDQTKIQYVIRAQLHVGA KQENWNTDIKTLKFDALSGSQEAEFTGYGKATLECQVQTAVDFGNSYI AEMEKESWIVDRQWAQDLTLPWQSGSGGVWREMHHLVEFEPPHAAT IRVLALGNQEGSLKTALTGAMRVTKDTNDNNLYKLHGGHVSCRVKLS ALTLKGTSYKMCTDKMSFVKNPTDTGHGTVVMQVKVPKGAPCRIPVI VADDLTAAINKGILVTVNPIASTNDDEVLIEVNPPFGDSYIIVGTGDSRL TYQWHKEGSSIGKLFTQTMKGAERLAVMGDAAWDFSSAGGFFTSVG KGIHTVFGSAFQGLFGGLSWITKVIMGAVLIWVGINTRNMTMSMSMIL VGVIMMFLSLGVGA YF_17D_JEVsp_ MWLVSLAIVTACAGAVTLVRKNRWLLLNVTSEDLGKTFSVGTGNCTT 112 SE_mutFL NILEAKYWCPDSMEYNCPNLSPREEPDDIDCWCYGVENVRVAYGKCD SAGRSRRSRRAIDLPTHENHGLKTRQEKWMTGRMGERQLQKIERWFV RNPFFAVTALTIAYLVGSNMTQRVVIALLVLAVGPAYSAIDLPTHENH GLKTRQEKWMTGRMGERQLQKIERWFVRNPFFAVTALTIAYLVGSN MTQRVVIALLVLAVGPAYSAHCIGITDRDFIEGVHGGTWVSATLEQDK CVTVMAPDKPSLDISLETVAIDRPAEVRKVCYNAVLTHVKINDKCPRE GEAHLAEENEGDNACKRTYSDRGRGNGCGRFGKGSIVACAKFTCAKS MSLFEVDQTKIQYVIRAQLHVGAKQENWNTDIKTLKFDALSGSQEVEF IGYGKATLECQVQTAVDFGNSYIAEMETESWIVDRQWAQDLTLPWQS GSGGVWREMHHLVEFEPPHAATIRVLALGNQEGSLKTALTGAMRVTK DTNDNNLYKLHGGHVSCRVKLSALTLKGTSYKICTDKMFFVKNPTDT GHGTVVMQVKVSKGAPCRIPVIVADDLTAAINKGILVTVNPIASTNDD EVLIEVNPPFGDSYIIVGRGDSRLTYQWHKEGSSIGKLFTQTMKGVERL AVMGDTAWDFSSAGGFFTSVGKGIHTVFGSAFQGLFGGLNWITKVIM GAVLIWVGINTRNMTMSMSMILVGVIMMFLSLGVGA YF_17D_IgGksp_ METPAQLLFLLLLWLPDTTGVTLVRKNRWLLLNVTSEDLGKTFSVGT 113 SE_mutFL GNCTTNILEAKYWCPDSMEYNCPNLSPREEPDDIDCWCYGVENVRVA YGKCDSAGRSRRSRRAIDLPTHENHGLKTRQEKWMTGRMGERQLQKI ERWFVRNPFFAVTALTIAYLVGSNMTQRVVIALLVLAVGPAYSAIDLP THENHGLKTRQEKWMTGRMGERQLQKIERWFVRNPFFAVTALTIAYL VGSNMTQRVVIALLVLAVGPAYSAHCIGITDRDFIEGVHGGTWVSATL EQDKCVTVMAPDKPSLDISLETVAIDRPAEVRKVCYNAVLTHVKINDK CPREGEAHLAEENEGDNACKRTYSDRGRGNGCGRFGKGSIVACAKFT CAKSMSLFEVDQTKIQYVIRAQLHVGAKQENWNTDIKTLKFDALSGS QEVEFIGYGKATLECQVQTAVDFGNSYIAEMETESWIVDRQWAQDLT LPWQSGSGGVWREMHHLVEFEPPHAATIRVLALGNQEGSLKTALTGA MRVTKDTNDNNLYKLHGGHVSCRVKLSALTLKGTSYKICTDKMFFVK NPTDTGHGTVVMQVKVSKGAPCRIPVIVADDLTAAINKGILVTVNPIAS TNDDEVLIEVNPPFGDSYIIVGRGDSRLTYQWHKEGSSIGKLFTQTMKG VERLAVMGDTAWDFSSAGGFFTSVGKGIHTVFGSAFQGLFGGLNWIT KVIMGAVLIWVGINTRNMTMSMSMILVGVIMMFLSLGVGA YF_17D_JEVsp_ MWLVSLAIVTACAGAVTLVRKNRWLLLNVTSEDLGKTFSVGTGNCTT 114 SE_no_polyNs_mut NILEAKYWCPDSMEYNCPNLSPREEPDDIDCWCYGVENVRVAYGKCD FL SAGRSRRSRRAIDLPTHENHGLKTRQEKWMTGRMGERQLQKIERWFV RNPFFAVTALTIAYLVGSNMTQRVVIALLVLAVGPAYSAIDLPTHENH GLKTRQEKWMTGRMGERQLQKIERWFVRNPFFAVTALTIAYLVGSN MTQRVVIALLVLAVGPAYSAHCIGITDRDFIEGVHGGTWVSATLEQDK CVTVMAPDKPSLDISLETVAIDRPAEVRKVCYNAVLTHVKINDKCPRE GEAHLAEENEGDNACKRTYSDRGRGNGCGRFGKGSIVACAKFTCAKS MSLFEVDQTKIQYVIRAQLHVGAKQENWNTDIKTLKFDALSGSQEVEF IGYGKATLECQVQTAVDFGNSYIAEMETESWIVDRQWAQDLTLPWQS GSGGVWREMHHLVEFEPPHAATIRVLALGNQEGSLKTALTGAMRVTK DTNDNNLYKLHGGHVSCRVKLSALTLKGTSYKICTDKMFFVKNPTDT GHGTVVMQVKVSKGAPCRIPVIVADDLTAAINKGILVTVNPIASTNDD EVLIEVNPPFGDSYIIVGRGDSRLTYQWHKEGSSIGKLFTQTMKGVERL AVMGDTAWDFSSAGGFFTSVGKGIHTVFGSAFQGLFGGLNWITKVIM GAVLIWVGINTRNMTMSMSMILVGVIMMFLSLGVGA YF_17D_JEVsp_ MWLVSLAIVTACAGAVTLVRKNRWLLLNVTSEDLGKTFSVGTGNCTT 115 SE_no_polyNs NILEAKYWCPDSMEYNCPNLSPREEPDDIDCWCYGVENVRVAYGKCD SAGRSRRSRRAIDLPTHENHGLKTRQEKWMTGRMGERQLQKIERWFV RNPFFAVTALTIAYLVGSNMTQRVVIALLVLAVGPAYSAIDLPTHENH GLKTRQEKWMTGRMGERQLQKIERWFVRNPFFAVTALTIAYLVGSN MTQRVVIALLVLAVGPAYSAHCIGITDRDFIEGVHGGTWVSATLEQDK CVTVMAPDKPSLDISLETVAIDRPAEVRKVCYNAVLTHVKINDKCPST GEAHLAEENEGDNACKRTYSDRGWGNGCGLFGKGSIVACAKFTCAKS MSLFEVDQTKIQYVIRAQLHVGAKQENWNTDIKTLKFDALSGSQEVEF IGYGKATLECQVQTAVDFGNSYIAEMETESWIVDRQWAQDLTLPWQS GSGGVWREMHHLVEFEPPHAATIRVLALGNQEGSLKTALTGAMRVTK DTNDNNLYKLHGGHVSCRVKLSALTLKGTSYKICTDKMFFVKNPTDT GHGTVVMQVKVSKGAPCRIPVIVADDLTAAINKGILVTVNPIASTNDD EVLIEVNPPFGDSYIIVGRGDSRLTYQWHKEGSSIGKLFTQTMKGVERL AVMGDTAWDFSSAGGFFTSVGKGIHTVFGSAFQGLFGGLNWITKVIM GAVLIWVGINTRNMTMSMSMILVGVIMMFLSLGVGA YF_17D_IgGksp_ METPAQLLFLLLLWLPDTTGVTLVRKNRWLLLNVTSEDLGKTFSVGT 116 SE_no_polyNs_mut GNCTTNILEAKYWCPDSMEYNCPNLSPREEPDDIDCWCYGVENVRVA FL YGKCDSAGRSRRSRRAIDLPTHENHGLKTRQEKWMTGRMGERQLQKI ERWFVRNPFFAVTALTIAYLVGSNMTQRVVIALLVLAVGPAYSAIDLP THENHGLKTRQEKWMTGRMGERQLQKIERWFVRNPFFAVTALTIAYL VGSNMTQRVVIALLVLAVGPAYSAHCIGITDRDFIEGVHGGTWVSATL EQDKCVTVMAPDKPSLDISLETVAIDRPAEVRKVCYNAVLTHVKINDK CPREGEAHLAEENEGDNACKRTYSDRGRGNGCGRFGKGSIVACAKFT CAKSMSLFEVDQTKIQYVIRAQLHVGAKQENWNTDIKTLKFDALSGS QEVEFIGYGKATLECQVQTAVDFGNSYIAEMETESWIVDRQWAQDLT LPWQSGSGGVWREMHHLVEFEPPHAATIRVLALGNQEGSLKTALTGA MRVTKDTNDNNLYKLHGGHVSCRVKLSALTLKGTSYKICTDKMFFVK NPTDTGHGTVVMQVKVSKGAPCRIPVIVADDLTAAINKGILVTVNPIAS TNDDEVLIEVNPPFGDSYIIVGRGDSRLTYQWHKEGSSIGKLFTQTMKG VERLAVMGDTAWDFSSAGGFFTSVGKGIHTVFGSAFQGLFGGLNWIT KVIMGAVLIWVGINTRNMTMSMSMILVGVIMMFLSLGVGA YF_17D_IgGksp_ METPAQLLFLLLLWLPDTTGVTLVRKNRWLLLNVTSEDLGKTFSVGT 117 SE_no_polyNs GNCTTNILEAKYWCPDSMEYNCPNLSPREEPDDIDCWCYGVENVRVA YGKCDSAGRSRRSRRAIDLPTHENHGLKTRQEKWMTGRMGERQLQKI ERWFVRNPFFAVTALTIAYLVGSNMTQRVVIALLVLAVGPAYSAIDLP THENHGLKTRQEKWMTGRMGERQLQKIERWFVRNPFFAVTALTIAYL VGSNMTQRVVIALLVLAVGPAYSAHCIGITDRDFIEGVHGGTWVSATL EQDKCVTVMAPDKPSLDISLETVAIDRPAEVRKVCYNAVLTHVKINDK CPSTGEAHLAEENEGDNACKRTYSDRGWGNGCGLFGKGSIVACAKFT CAKSMSLFEVDQTKIQYVIRAQLHVGAKQENWNTDIKTLKFDALSGS QEVEFIGYGKATLECQVQTAVDFGNSYIAEMETESWIVDRQWAQDLT LPWQSGSGGVWREMHHLVEFEPPHAATIRVLALGNQEGSLKTALTGA MRVTKDTNDNNLYKLHGGHVSCRVKLSALTLKGTSYKICTDKMFFVK NPTDTGHGTVVMQVKVSKGAPCRIPVIVADDLTAAINKGILVTVNPIAS TNDDEVLIEVNPPFGDSYIIVGRGDSRLTYQWHKEGSSIGKLFTQTMKG VERLAVMGDTAWDFSSAGGFFTSVGKGIHTVFGSAFQGLFGGLNWIT KVIMGAVLIWVGINTRNMTMSMSMILVGVIMMFLSLGVGA

Underlined sequence corresponds to a signal peptide, which may be omitted from each sequence. Thus, any RNA vaccine provided herein may encode an antigen represented by a sequence of Table 22, with or without the underlined signal peptide.

TABLE 23 ZIKV mRNA Vaccine Immunogenicity Study Study design BALB/C Immunization Group Vaccine N Dose Route Prime Boost Endpoint 1 Zika prME 5 10 ug IM Day 0 Day 21 Terminal bleeds on Day vaccine 41. Anti Zika 2 Zika prME 5 10 ug IM Day 0 NA neutralizing IgG titer. vaccine 3 Zika prME 5  2 ug IM Day 0 Day 21 vaccine 4 Zika prME 5  2 ug IM Day 0 NA vaccine 5 PBS 5 NA IM Day 0 Day 21

TABLE 24 Example 24 Test Conditions Study design AG129 Immunization Group Vaccine n Dose Route Prime Boost Challenge Endpoint 1 Zika prME 8 10 ug IM Day 0 Day 21 Day 42 Monitor for vaccine survival 2 Zika prME 8 10 ug IM Day 0 NA and weight vaccine loss. Viral 3 Zika prME 8  2 ug IM Day 0 Day 21 load at Day 5 vaccine 4 Zika prME 8  2 ug IM Day 0 NA vaccine 5 PBS 8 NA IM Day 0 Day 21

TABLE 25 ZIKV Nucleic Acid Sequences SEQ ID Description Sequence NO: Zika virus strain ATGAAAAACCCAAAGAAGAAATCCGGAGGATTCCGGATTGTCAATAT 118 MR 766 GCTAAAACGCGGAGTAGCCCGTGTAAACCCCTTGGGAGGTTTGAAGAG polyprotein gene, GCTGCCAGCCGGACTTCTGCTGGGTCATGGACCCATCAGAATGGTTTT complete cds GGCGATATTAGCCTTTTTGAGATTCACAGCAATCAAGCCATCACTGGG GenBank CCTCATCAACAGATGGGGTACCGTGGGGAAAAAAGAGGCTATGGAAA Accession: TAATAAAAAAATTTAAGAAAGATCTTGCTGCCATGTTGAGAATAATCA DQ859059 ATGCTAGGAAGGAGAGGAAGAGACGTGGCGCAGACACCAGCATCGGA ATCGTTGGCCTCCTGTTGACTACAGCCATGGCAGCAGAGATCACTAGA CGTGGGAGTGCATACTACATGTACTTGGATAGGAGCGATGCAGGGAA GGCCATTTCTTTCGCTACCACATTGGGGGTGAACAAATGCCATGTGCA GATCATGGACCTCGGGCACATGTGTGACGCCACCATGAGCTATGAATG CCCTATGCTGGACGAGGGGGTGGAACCAGATGACGTCGATTGCTGGTG CAACACGACATCAACTTGGGTTGTGTACGGAACCTGTCATCATAAAAA AGGTGAAGCACGGCGATCTAGAAGAGCCGTCACGCTCCCATCTCACTC CACAAGGAAATTGCAAACGCGGTCGCAGACTTGGCTAGAATCAAGAG AATACACAAAGCACCTGATCAAGGTTGAAAATTGGATATTCAGGAACC CTGGTTTTACGCTAGTGGCTGTCGCCATCGCCTGGCTTTTGGGAAGCTC GACGAGCCAAAAAGTCATATACTTGGTCATGATACTGCTGATTGCCCC GGCATACAGTATCAGGTGCATAGGAGTCAGCAATAGAGACTTCGTGGA GGGCATGTCAGGTGGGACCTGGGTTGACGTTGTCCTGGAACATGGAGG CTGCGTCACCGTGATGGCACAGGACAAGCCAACAGTTGACATAGAGCT GGTCACAACAACGGTTAGTAACATGGCCGAGGTGAGATCCTATTGTTA CGAGGCATCAATATCGGACATGGCTTCGGACAGTCGCTGCCCAACACA AGGTGAAGCCTACCTTGACAAGCAATCAGACACTCAATATGTTTGCAA AAGAACATTGGTGGACAGAGGTTGGGGAAATGGGTGTGGACTCTTTGG CAAAGGGAGTTTGGTGACATGTGCTAAGTTCACGTGCTCCAAGAAGAT GACTGGGAAGAGCATTCAGCCGGAGAACCTGGAGTATCGGATAATGC TATCAGTGCATGGCTCCCAGCACAGTGGGATGATTGTTAATGATGAAA ACAGAGCGAAGGTCGAGGTTACGCCCAATTCACCAAGAGCAGAAGCA ACCCTGGGAGGCTTTGGAAGCTTAGGACTTGATTGTGAACCAAGGACA GGCCTTGACTTTTCAGATCTGTATTACCTAACCATGAATAACAAGCATT GGTTGGTGCACAAAGAGTGGTTTCATGACATCCCATTGCCCTGGCATG CTGGGGCAGACACTGGAACTCCACATTGGAACAACAAGGAGGCATTA GTGGAATTCAAGGACGCCCACGCCAAGAGGCAAACCGTCGTGGTTTTG GGGAGCCAGGAAGGAGCCGTCCACACGGCTCTTGCTGGAGCTCTAGA GGCTGAGATGGATGGTGCAAAGGGAAGGCTATTCTCTGGCCACTTGAA ATGTCGCTTAAAAATGGACAAGCTTAGATTGAAGGGCGTGTCATATTC CTTGTGCACCGCGGCATTCACATTCACCAAGGTCCCGGCTGAAACACT ACATGGAACAGTCACAGTGGAGGTGCAGTATGCAGGGACAGATGGAC CCTGCAAGGTCCCAGCCCAGATGGCGGTGGACATGCAGACCTTGACCC CAGTCGGAAGGCTGATAACCGCCAACCCCGTGATTACTGAAAGCACTG AGAATTCAAAGATGATGTTGGAGCTCGACCCACCATTTGGGGATTCTT ACATTGTCATAGGAGTTGGGGATAAGAAAATCACCCATCACTGGCATA GGAGTGGCAGCACCATTGGAAAAGCATTTGAAGCCACTGTGAGAGGC GCTAAGAGAATGGCAGTCCTGGGGGACACAGCTTGGGACTTTGGATCA GTCGGAGGTGTGTTTAACTCATTGGGCAAGGGCATTCATCAGATTTTTG GAGCAGCTTTCAAATCACTGTTTGGAGGAATGTCCTGGTTCTCACAGA TCCTCATAGGCACTCTGCTGGTGTGGTTAGGTCTGAACACAAAGAATG GGTCTATCTCCCTCACATGCTTAGCCCTGGGGGGAGTGATGATCTTCCT CTCCACGGCTGTTTCTGCTGACGTGGGGTGCTCGGTGGACTTCTCAAAA AAAGAAACGAGATGTGGCACGGGGGTGTTCGTCTACAATGACGTTGA AGCCTGGAGGGACCGGTACAAGTACCATCCTGACTCCCCTCGTAGACT GGCAGCAGCCGTTAAGCAAGCTTGGGAAGAGGGGATTTGTGGGATCTC CTCTGTTTCTAGAATGGAAAACATAATGTGGAAATCAGTGGAAGGAGA GCTCAATGCAATCCTAGAGGAGAATGGAGTCCAACTGACAGTTGTTGT GGGATCTGTAAAAAACCCCATGTGGAGAGGCCCACAAAGATTGCCAG TGCCTGTGAATGAGCTGCCCCATGGCTGGAAAGCCTGGGGGAAATCGT ACTTTGTTAGGGCGGCAAAGACCAACAACAGTTTTGTTGTCGACGGTG ACACATTGAAGGAATGTCCGCTCAAGCACAGAGCATGGAACAGCTTCC TCGTGGAGGATCACGGGTTTGGGGTCTTCCACACCAGTGTTTGGCTTA AGGTTAGAGAAGATTACTCACTGGAGTGTGACCCAGCCGTCATAGGAA CAGCTGTTAAGGGAAAGGAGGCCGCGCACAGTGATCTAGGCTATTGG ATTGAAAGTGAAAAGAATGACACATGGAGGCTGAAGAGGGCTCATTT GATTGAGATGAAAACATGTGAGTGGCCAAAGTCTCACACACTGTGGAC AGATGGAGTGGAAGAAAGTGATCTGATCATACCCAAGTCTTTAGCTGG TCCACTCAGCCACCACAACACCAGAGAGGGTTACAGAACTCAAGTGA AAGGGCCATGGCATAGTGAGGAGCTTGAAATCCGATTTGAGGAATGTC CAGGTACCAAGGTTCATGTGGAGGAGACATGCGGAACGAGAGGACCA TCTCTGAGATCAACCACTGCAAGCGGAAGGGTCATTGAGGAATGGTGC TGTAGGGAATGCACAATGCCCCCACTATCGTTCCGAGCAAAAGATGGC TGCTGGTATGGAATGGAGATAAGGCCTAGGAAAGAACCAGAGAGCAA CTTAGTGAGGTCAATGGTGACAGCGGGATCAACCGATCATATGGATCA TTTTTCTCTTGGAGTGCTTGTGATTCTACTCATGGTGCAGGAAGGGTTG AAGAAGAGAATGACCACAAAGATCATCATGAGCACATCAATGGCAGT GCTGGTGGCCATGATCTTGGGAGGATTCTCAATGAGTGACCTGGCTAA GCTTGTGATCCTGATGGGGGCCACTTTCGCAGAAATGAACACTGGAGG AGACGTAGCTCACTTGGCATTAGTAGCGGCATTTAAAGTCAGACCAGC CTTGCTGGTCTCATTTATCTTCAGAGCCAACTGGACACCTCGTGAGAGC ATGCTGCTAGCCTTGGCTTCGTGTCTTCTGCAAACTGCGATCTCCGCTC TTGAAGGCGACTTGATGGTCCTCGTTAATGGATTTGCTTTGGCCTGGTT GGCAATACGTGCAATGGCCGTGCCACGCACTGACAACATCGCTCTAGC AATTCTGGCTGCTCTAACACCACTAGCCCGAGGCACACTGCTCGTGGC ATGGAGAGCGGGCCTCGCCACTTGTGGAGGGTTCATGCTCCTCTCCCT GAAAGGGAAAGGTAGTGTGAAGAAGAACCTGCCATTCGTCGCGGCCT TGGGATTGACCGCTGTGAGAATAGTGGACCCCATTAATGTGGTGGGAC TACTGTTACTCACAAGGAGTGGGAAGCGGAGCTGGCCCCCTAGTGAAG TGCTCACTGCTGTCGGCCTGATATGTGCATTGGCCGGAGGGTTTGCCA AGGCAGACATAGAGATGGCTGGGCCCATGGCGGCAGTGGGCCTGCTA ATTGTCAGTTATGTGGTCTCGGGAAAGAGTGTAGATATGTACATTGAA AGAGCAGGTGACATCACATGGGAGAAAGACGCGGAAGTCACTGGAAA CAGTCCTCGGCTTGACGTGGCACTAGATGAGAGTGGTGATTTCTCTCTG GTGGAGGAAGATGGTCCACCCATGAGAGAGATCATACTTAAGGTGGTC TTGATGGCCATCTGTGGCATGAACCCAATAGCCATACCTTTTGCTGCAG GAGCGTGGTATGTGTATGTGAAGACTGGGAAAAGGAGTGGTGCCCTCT GGGACGTGCCTGCTCCGAAAGAAGTGAAAAAAGGAGAGACCACAGAT GGAGTGTACAGAGTGATGACTCGCAGACTGCTGGGTTCAACACAAGTT GGAGTGGGAGTCATGCAGGAGGGAGTCTTCCACACCATGTGGCACGTC ACAAAAGGGGCCGCATTGAGGAGCGGTGAAGGGAGACTTGATCCATA CTGGGGGGATGTCAAGCAGGACTTGGTGTCATATTGTGGGCCTTGGAA GCTGGACGCAGCTTGGGACGGAGTTAGTGAGGTGCAGCTTCTGGCCGT ACCCCCTGGAGAGAGAGCCAGAAACATTCAGACTCTGCCTGGAATATT TAAGACAAAGGATGGGGACATCGGAGCAGTTGCTTTGGACTATCCTGC AGGAACCTCAGGATCTCCGATCCTAGACAAATGCGGGAGAGTGATAG GACTCTATGGCAATGGGGTTGTGATCAAGAACGGAAGCTATGTTAGTG CTATAACCCAGGGAAAGAGGGAGGAGGAGACTCCGGTTGAGTGTTTT GAACCCTCGATGCTGAAGAAGAAGCAGCTAACTGTCCTGGACCTGCAT CCAGGGGCTGGGAAAACCAGGAGAGTTCTTCCTGAAATAGTCCGTGAA GCTATAAAGAAGAGACTCCGCACGGTGATCTTGGCACCAACCAGGGTC GTCGCTGCTGAGATGGAGGAAGCCCTGAGAGGACTTCCGGTGCGTTAC ATGACAACAGCAGTCAAGGTCACCCATTCTGGGACAGAAATCGTTGAT TTGATGTGCCATGCCACCTTCACTTCACGCCTACTACAACCCATTAGAG TCCCTAATTACAACCTCTACATCATGGATGAAGCCCATTTCACAGACCC CTCAAGCATAGCTGCAAGAGGATATATATCAACAAGGGTTGAGATGG GCGAGGCAGCAGCCATCTTTATGACTGCCACACCACCAGGAACCCGCG ATGCGTTTCCAGATTCCAACTCACCAATCATGGACACAGAAGTGGAAG TCCCAGAGAGAGCCTGGAGCTCAGGCTTTGATTGGGTGACGGACCATT CTGGGAAAACAGTTTGGTTCGTTCCAAGCGTGAGGAATGGAAATGAAA TCGCAGCCTGTCTGACAAAGGCTGGAAAGCGGGTTATACAGCTTAGTA GGAAAACTTTTGAGACAGAGTTTCAGAAAACAAAAAATCAAGAGTGG GACTTTGTCATAACAACTGACATCTCAGAGATGGGTGCCAACTTCAAG GCTGACCGGGTTATAGATTCCAGGAGATGCCTAAAGCCAGTTATACTT GATGGTGAGAGAGTCATCTTGGCTGGGCCCATGCCTGTCACGCATGCT AGCGCTGCTCAGAGGAGAGGACGTATAGGCAGGAACCCCAACAAGCC TGGAGATGAGTACATGTATGGAGGTGGGTGTGCGGAGACTGATGAAG ACCATGCACATTGGCTTGAAGCAAGAATGCTTCTTGACAACATTTACC TCCAGGATGGCCTCATAGCCTCGCTCTATCGACCTGAGGCCGACAAGG TAGCCGCCATTGAGGGAGAGTTTAAGCTGAGGACAGAGCAAAGGAAG ACCTTTGTGGAACTCATGAAGAGAGGAGATCTTCCCGTTTGGTTGGCC TACCAGGTTGCATCTGCCGGAATAACTTATACAGACAGAAGATGGTGT TTTGATGGCACAACCAACAACACCATAATGGAAGACAGTGTACCAGCA GAGGTGTGGACCAAGTATGGAGAGAAGAGAGTGCTCAAACCAAGATG GATGGACGCCAGGGTCTGCTCAGATCATGCGGCCCTGAAGTCGTTCAA AGAATTCGCCGCTGGGAAAAGAGGAGCGGCTTTGGGAGTAATGGAGG CCCTGGGAACATTACCAGGACACATGACAGAGAGGTTTCAGGAAGCC ATTGATAACCTCGCTGTGCTCATGCGAGCAGAGACTGGAAGCAGGCCC TACAAGGCAGCGGCAGCCCAATTGCCGGAGACCCTAGAGACCATCAT GCTTTTAGGCCTGCTGGGAACAGTATCGCTGGGGATCTTTTTTGTCTTG ATGAGGAACAAGGGCATCGGGAAGATGGGCTTTGAAATGGTAACCCT TGGGGCCAGCGCATGGCTCATGTGGCTCTCAGAAATCGAACCAGCCAG AATTGCATGTGTCCTTATTGTTGTGTTTTTATTACTGGTGGTGCTAATAC CAGAGCCAGAGAAGCAAAGATCCCCCCAGGACAATCAGATGGCAATC ATTATTATGGTGGCAGTGGGCCTTTTGGGGTTGATAACTGCAAATGAA CTTGGATGGCTGGAGAGAACAAAAAATGACATAGCTCATCTGATGGG AAAGAGAGAAGAGGGAACAACCGTGGGATTCTCAATGGACATCGATC TGCGACCAGCCTCCGCATGGGCTATTTATGCCGCATTGACAACCCTCAT CACCCCAGCCGTCCAGCACGCGGTAACTACCTCGTACAACAACTACTC CTTAATGGCGATGGCCACACAAGCTGGAGTGCTGTTTGGCATGGGCAA AGGGATGCCATTTTATGCATGGGACTTAGGAGTCCCGTTGCTAATGAT GGGCTGCTACTCACAACTAACACCCCTGACCCTGATAGTAGCCATCAT TTTGCTTGTGGCACATTACATGTACTTGATCCCAGGCCTACAGGCAGCA GCAGCACGCGCTGCCCAGAAGAGAACAGCAGCCGGCATCATGAAGAA TCCCGTTGTGGATGGAATAGTGGTAACTGACATTGACACAATGACAAT TGACCCCCAAGTGGAGAAGAAGATGGGACAAGTGCTACTTATAGCAG TGGCTGTCTCCAGTGCTGTGTTGCTGCGGACCGCTTGGGGATGGGGGG AGGCTGGAGCTTTGATCACAGCAGCAACTTCCACCCTGTGGGAAGGCT CCCCAAACAAATACTGGAACTCCTCCACAGCCACCTCACTGTGCAACA TCTTCAGAGGAAGTTACTTGGCAGGAGCTTCCCTTATTTACACAGTGAC AAGAAATGCCGGCCTGGTTAAGAGACGTGGAGGTGGAACGGGAGAAA CTCTGGGAGAGAAGTGGAAAGCCCGCCTGAATCAGATGTCGGCCTTGG AGTTCTACTCTTACAAAAAGTCAGGCATCACTGAAGTATGTAGAGAGG AGGCTCGCCGCGCCCTCAAGGATGGAGTGGCCACAGGAGGACATGCT GTATCCCGAGGAAGCGCAAAACTCAGATGGTTGGTGGAGAGAGGATA TCTGCAGCCCTATGGAAAGGTTGTTGATCTCGGATGCGGCAGAGGGGG CTGGAGTTATTATGCCGCCACCATCCGCAAAGTGCAGGAGGTGAGAGG ATACACAAAGGGAGGTCCCGGTCATGAAGAGCCCATGCTGGTGCAAA GCTATGGGTGGAACATAATTCGTCTCAAGAGTGGAGTGGACGTCTTCC ACATGGCGGCTGAGTCGTGTGACACTTTGCTGTGTGACATAGGTGAGT CATCATCCAGTCCTGAAGTGGAGGAGACGCGAACACTCAGAGTGCTCT CCATGGTGGGGGACTGGCTTGAGAAGAGACCAGGGGCCTTCTGCATAA AGGTGTTATGCCCATACACCAGCACCATGATGGAGACCATGGAGCGAC TGCAACGTAGGTATGGGGGAGGACTAGTCAGAGTGCCACTGTCCCGCA ATTCTACACATGAGATGTATTGGGTCTCTGGAGCAAAAAGTAACATCA TAAAAAGTGTGTCCACCACAAGTCAGCTCCTCCTGGGACGCATGGATG GGCCCAGGAGGCCAGTGAAGTATGAGGAGGATGTGAACCTCGGCTCA GGCACACGAGCTGTGGCAAGCTGTGCTGAGGCTCCCAACATGAAGGTC ATTGGTAGGCGCATTGAGAGAATCCGTAGTGAACATGCAGAAACATG GTTCTTTGATGAAAACCATCCATACAGGACATGGGCCTACCACGGGAG CTACGAAGCCCCCACGCAAGGGTCAGCATCTTCCCTCGTGAATGGGGT TGTTAGACTCCTGTCAAAGCCCTGGGATGTGGTGACTGGAGTTACAGG AATAGCTATGACTGACACCACACCGTACGGCCAACAAAGAGTCTTCAA AGAAAAAGTGGACACCAGGGTGCCAGACCCTCAAGAAGGTACTCGCC AGGTAATGAACATGGTCGCTTCCTGGCTGTGGAAGGAGCTGGGAAAAC GTAAGCGGCCACGTGTCTGCACCAAAGAAGAGTTCATCAACAAGGTGC GCAGCAATGCAGCACTGGGAGCAATATTTGAAGAGGAAAAAGAATGG AAGACGGCTGTGGAAGCTGTGAATGATCCAAGGTTTTGGGCCCTAGTG GATAAGGAAAGAGAACACCACCTGAGAGGAGAGTGCCATAGTTGTGT GTACAACATGATGGGAAAAAGAGAAAAGAAGCAAGGGGAATTCGGG AAAGCAAAAGGCAGTCGCGCCATCTGGTACATGTGGTTGGGAGCCAG ATTCTTGGAGTTTGAAGCCCTTGGATTCTTGAACGAGGACCATTGGAT GGGAAGAGAAAACTCAGGAGGTGGTGTCGAAGGGTTGGGACTGCAAA GACTTGGATACGTTCTAGAAGAAATGAGCCGGGCACCAGGAGGAAAG ATGTATGCAGATGACACCGCTGGCTGGGACACCCGCATTAGCAAGTTT GATTTGGAGAATGAAGCCTTGATTACTAACCAAATGGATGAAGGGCAC AGAACTCTGGCGTTGGCCGTGATTAAGTACACATACCAAAACAAAGTG GTGAAGGTCCTCAGACCAGCTGAAGGAGGAAAAACAGTCATGGACAT CATTTCAAGACAAGACCAGAGGGGGAGCGGACAAGTTGTCACTTATGC TCTCAACACATTTACCAACTTGGTGGTGCAGCTCATCCGGAACATGGA GGCTGAGGAAGTGTTAGAGATGCAAGACTTATGGCTGTTGAGGAAGCC AGAGAAAGTAACCAGATGGCTGCAGAGTAGCGGATGGGACAGACTCA AACGAATGGCAGTCAGTGGTGATGACTGTGTTGTAAAGCCAATTGATG ACAGGTTTGCACACGCCCTCAGGTTCTTGAATGATATGGGGAAAGTTA GGAAAGACACACAGGAATGGAAACCCTCAACTGGATGGAGCAACTGG GAAGAAGTCCCGTTCTGCTCCCACCACTTTAACAAGCTGCACCTCAAA GACGGGAGATCCATTGTGGTCCCTTGCCGCCACCAAGATGAACTGATT GGCCGGGCTCGCGTTTCGCCGGGGGCAGGATGGAGCATCCGGGAGAC TGCCTGTCTTGCAAAATCATATGCACAGATGTGGCAGCTTCTTTATTTC CACAGAAGAGACCTCCGACTGATGGCCAATGCCATTTGCTCGGCCGTG CCAGTTGACTGGGTCCCAACTGGGAGAACTACCTGGTCAATCCATGGA AAGGGAGAATGGATGACTACTGAGGACATGCTCATGGTGTGGAATAG AGTGTGGATTGAGGAGAATGATCACATGGAGGACAAGACCCCTGTAA CAAAATGGACAGACATTCCCTATTTGGGAAAAAGGGAGGACTTATGGT GTGGATCCCTTATAGGACACAGACCTCGCACCACTTGGGCTGAGAACA TCAAAGACACAGTCAGCATGGTGCGCAGAATCATAGGTGATGAAGAA AAGTACATGGACTACCTATCCACTCAAGTTCGCTACTTGGGTGAGGAA GGGTCTACACCTGGAGTGCTGTAA IgE HC signal TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGG 119 peptide_prM-E #1 AAATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCAT (Brazil_isolate_ GGACTGGACCTGGATCCTGTTCCTGGTGGCCGCTGCCACAAGAGTGCA ZikaSPH2015, CAGCGTGGAAGTGACCAGACGGGGCAGCGCCTACTACATGTACCTGG Sequence, NT (5′ ACAGAAGCGACGCCGGCGAGGCCATCAGCTTTCCAACCACCCTGGGCA UTR, ORF, 3′ TGAACAAGTGCTACATCCAGATCATGGACCTGGGCCACATGTGCGACG UTR) CCACCATGAGCTACGAGTGCCCCATGCTGGACGAGGGCGTGGAACCCG ACGATGTGGACTGCTGGTGCAACACCACCAGCACCTGGGTGGTGTACG GCACCTGTCACCACAAGAAGGGCGAAGCCAGACGGTCCAGACGGGCC GTGACACTGCCTAGCCACAGCACCAGAAAGCTGCAGACCCGGTCCCAG ACCTGGCTGGAAAGCAGAGAGTACACCAAGCACCTGATCCGGGTGGA AAACTGGATCTTCCGGAACCCCGGCTTTGCCCTGGCTGCCGCTGCTATT GCTTGGCTGCTGGGCAGCAGCACCTCCCAGAAAGTGATCTACCTCGTG ATGATCCTGCTGATCGCCCCTGCCTACAGCATCCGGTGTATCGGCGTGT CCAACCGGGACTTCGTGGAAGGCATGAGCGGCGGCACATGGGTGGAC GTGGTGCTGGAACATGGCGGCTGCGTGACAGTGATGGCCCAGGATAA GCCCGCCGTGGACATCGAGCTCGTGACCACCACCGTGTCCAATATGGC CGAAGTGCGGAGCTACTGCTACGAGGCCAGCATCAGCGACATGGCCA GCGACAGCAGATGCCCTACACAGGGCGAGGCCTACCTGGATAAGCAG TCCGACACCCAGTACGTGTGCAAGCGGACCCTGGTGGATAGAGGCTGG GGCAATGGCTGCGGCCTGTTTGGCAAGGGCAGCCTCGTGACCTGCGCC AAGTTCGCCTGCAGCAAGAAGATGACCGGCAAGAGCATCCAGCCCGA GAACCTGGAATACCGGATCATGCTGAGCGTGCACGGCTCCCAGCACAG CGGCATGATCGTGAACGACACCGGCCACGAGACAGACGAGAACCGGG CCAAGGTGGAAATCACCCCCAACAGCCCTAGAGCCGAGGCCACACTG GGCGGCTTTGGATCTCTGGGCCTGGACTGCGAGCCTAGAACCGGCCTG GATTTCAGCGACCTGTACTACCTGACCATGAACAACAAGCACTGGCTG GTGCACAAAGAGTGGTTCCACGACATCCCCCTGCCCTGGCATGCTGGC GCTGATACAGGCACCCCCCACTGGAACAACAAAGAGGCTCTGGTGGA ATTCAAGGACGCCCACGCCAAGCGGCAGACCGTGGTGGTGCTGGGATC TCAGGAAGGCGCCGTGCATACAGCTCTGGCTGGCGCCCTGGAAGCCGA AATGGATGGCGCCAAAGGCAGACTGTCCTCCGGCCACCTGAAGTGCCG GCTGAAGATGGACAAGCTGCGGCTGAAGGGCGTGTCCTACAGCCTGTG TACCGCCGCCTTCACCTTCACCAAGATCCCCGCCGAGACACTGCACGG CACCGTGACTGTGGAAGTGCAGTACGCCGGCACCGACGGCCCTTGTAA AGTGCCTGCTCAGATGGCCGTGGATATGCAGACCCTGACCCCCGTGGG CAGACTGATCACCGCCAACCCTGTGATCACCGAGAGCACCGAGAACA GCAAGATGATGCTGGAACTGGACCCCCCCTTCGGCGACTCCTACATCG TGATCGGCGTGGGAGAGAAGAAGATCACCCACCACTGGCACAGATCC GGCAGCACCATCGGCAAGGCCTTTGAGGCTACAGTGCGGGGAGCCAA GAGAATGGCCGTGCTGGGCGATACCGCCTGGGATTTTGGCTCTGTGGG CGGAGCCCTGAACAGCCTGGGAAAGGGCATCCACCAGATCTTCGGCGC TGCCTTCAAGAGCCTGTTCGGCGGCATGAGCTGGTTCAGCCAGATCCT GATCGGCACCCTGCTCGTGTGGCTGGGCCTGAACACCAAGAACGGCAG CATCTCCCTGACCTGCCTGGCTCTGGGCGGCGTGCTGATCTTTCTGAGC ACAGCCGTGTCCGCCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTT CTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGT ACCCCCGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC IgE HC signal ATGGACTGGACCTGGATCCTGTTCCTGGTGGCCGCTGCCACAAGAGTG 120 peptide_prM-E #1 CACAGCGTGGAAGTGACCAGACGGGGCAGCGCCTACTACATGTACCTG (Brazil_isolate_ GACAGAAGCGACGCCGGCGAGGCCATCAGCTTTCCAACCACCCTGGGC ZikaSPH2015), ORF ATGAACAAGTGCTACATCCAGATCATGGACCTGGGCCACATGTGCGAC Sequence, NT GCCACCATGAGCTACGAGTGCCCCATGCTGGACGAGGGCGTGGAACCC GACGATGTGGACTGCTGGTGCAACACCACCAGCACCTGGGTGGTGTAC GGCACCTGTCACCACAAGAAGGGCGAAGCCAGACGGTCCAGACGGGC CGTGACACTGCCTAGCCACAGCACCAGAAAGCTGCAGACCCGGTCCCA GACCTGGCTGGAAAGCAGAGAGTACACCAAGCACCTGATCCGGGTGG AAAACTGGATCTTCCGGAACCCCGGCTTTGCCCTGGCTGCCGCTGCTAT TGCTTGGCTGCTGGGCAGCAGCACCTCCCAGAAAGTGATCTACCTCGT GATGATCCTGCTGATCGCCCCTGCCTACAGCATCCGGTGTATCGGCGT GTCCAACCGGGACTTCGTGGAAGGCATGAGCGGCGGCACATGGGTGG ACGTGGTGCTGGAACATGGCGGCTGCGTGACAGTGATGGCCCAGGATA AGCCCGCCGTGGACATCGAGCTCGTGACCACCACCGTGTCCAATATGG CCGAAGTGCGGAGCTACTGCTACGAGGCCAGCATCAGCGACATGGCC AGCGACAGCAGATGCCCTACACAGGGCGAGGCCTACCTGGATAAGCA GTCCGACACCCAGTACGTGTGCAAGCGGACCCTGGTGGATAGAGGCTG GGGCAATGGCTGCGGCCTGTTTGGCAAGGGCAGCCTCGTGACCTGCGC CAAGTTCGCCTGCAGCAAGAAGATGACCGGCAAGAGCATCCAGCCCG AGAACCTGGAATACCGGATCATGCTGAGCGTGCACGGCTCCCAGCACA GCGGCATGATCGTGAACGACACCGGCCACGAGACAGACGAGAACCGG GCCAAGGTGGAAATCACCCCCAACAGCCCTAGAGCCGAGGCCACACT GGGCGGCTTTGGATCTCTGGGCCTGGACTGCGAGCCTAGAACCGGCCT GGATTTCAGCGACCTGTACTACCTGACCATGAACAACAAGCACTGGCT GGTGCACAAAGAGTGGTTCCACGACATCCCCCTGCCCTGGCATGCTGG CGCTGATACAGGCACCCCCCACTGGAACAACAAAGAGGCTCTGGTGG AATTCAAGGACGCCCACGCCAAGCGGCAGACCGTGGTGGTGCTGGGA TCTCAGGAAGGCGCCGTGCATACAGCTCTGGCTGGCGCCCTGGAAGCC GAAATGGATGGCGCCAAAGGCAGACTGTCCTCCGGCCACCTGAAGTGC CGGCTGAAGATGGACAAGCTGCGGCTGAAGGGCGTGTCCTACAGCCTG TGTACCGCCGCCTTCACCTTCACCAAGATCCCCGCCGAGACACTGCAC GGCACCGTGACTGTGGAAGTGCAGTACGCCGGCACCGACGGCCCTTGT AAAGTGCCTGCTCAGATGGCCGTGGATATGCAGACCCTGACCCCCGTG GGCAGACTGATCACCGCCAACCCTGTGATCACCGAGAGCACCGAGAA CAGCAAGATGATGCTGGAACTGGACCCCCCCTTCGGCGACTCCTACAT CGTGATCGGCGTGGGAGAGAAGAAGATCACCCACCACTGGCACAGAT CCGGCAGCACCATCGGCAAGGCCTTTGAGGCTACAGTGCGGGGAGCC AAGAGAATGGCCGTGCTGGGCGATACCGCCTGGGATTTTGGCTCTGTG GGCGGAGCCCTGAACAGCCTGGGAAAGGGCATCCACCAGATCTTCGG CGCTGCCTTCAAGAGCCTGTTCGGCGGCATGAGCTGGTTCAGCCAGAT CCTGATCGGCACCCTGCTCGTGTGGCTGGGCCTGAACACCAAGAACGG CAGCATCTCCCTGACCTGCCTGGCTCTGGGCGGCGTGCTGATCTTTCTG AGCACAGCCGTGTCCGCC IgE HC signal G*GGGAAATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCC 121 peptide_prM-E #1 ACCATGGACTGGACCTGGATCCTGTTCCTGGTGGCCGCTGCCACAAGA (Brazil_isolate_ GTGCACAGCGTGGAAGTGACCAGACGGGGCAGCGCCTACTACATGTA ZikaSPH2015), mRNA CCTGGACAGAAGCGACGCCGGCGAGGCCATCAGCTTTCCAACCACCCT Sequence (T100 GGGCATGAACAAGTGCTACATCCAGATCATGGACCTGGGCCACATGTG tail) CGACGCCACCATGAGCTACGAGTGCCCCATGCTGGACGAGGGCGTGG AACCCGACGATGTGGACTGCTGGTGCAACACCACCAGCACCTGGGTGG TGTACGGCACCTGTCACCACAAGAAGGGCGAAGCCAGACGGTCCAGA CGGGCCGTGACACTGCCTAGCCACAGCACCAGAAAGCTGCAGACCCG GTCCCAGACCTGGCTGGAAAGCAGAGAGTACACCAAGCACCTGATCC GGGTGGAAAACTGGATCTTCCGGAACCCCGGCTTTGCCCTGGCTGCCG CTGCTATTGCTTGGCTGCTGGGCAGCAGCACCTCCCAGAAAGTGATCT ACCTCGTGATGATCCTGCTGATCGCCCCTGCCTACAGCATCCGGTGTAT CGGCGTGTCCAACCGGGACTTCGTGGAAGGCATGAGCGGCGGCACAT GGGTGGACGTGGTGCTGGAACATGGCGGCTGCGTGACAGTGATGGCCC AGGATAAGCCCGCCGTGGACATCGAGCTCGTGACCACCACCGTGTCCA ATATGGCCGAAGTGCGGAGCTACTGCTACGAGGCCAGCATCAGCGAC ATGGCCAGCGACAGCAGATGCCCTACACAGGGCGAGGCCTACCTGGA TAAGCAGTCCGACACCCAGTACGTGTGCAAGCGGACCCTGGTGGATAG AGGCTGGGGCAATGGCTGCGGCCTGTTTGGCAAGGGCAGCCTCGTGAC CTGCGCCAAGTTCGCCTGCAGCAAGAAGATGACCGGCAAGAGCATCC AGCCCGAGAACCTGGAATACCGGATCATGCTGAGCGTGCACGGCTCCC AGCACAGCGGCATGATCGTGAACGACACCGGCCACGAGACAGACGAG AACCGGGCCAAGGTGGAAATCACCCCCAACAGCCCTAGAGCCGAGGC CACACTGGGCGGCTTTGGATCTCTGGGCCTGGACTGCGAGCCTAGAAC CGGCCTGGATTTCAGCGACCTGTACTACCTGACCATGAACAACAAGCA CTGGCTGGTGCACAAAGAGTGGTTCCACGACATCCCCCTGCCCTGGCA TGCTGGCGCTGATACAGGCACCCCCCACTGGAACAACAAAGAGGCTCT GGTGGAATTCAAGGACGCCCACGCCAAGCGGCAGACCGTGGTGGTGC TGGGATCTCAGGAAGGCGCCGTGCATACAGCTCTGGCTGGCGCCCTGG AAGCCGAAATGGATGGCGCCAAAGGCAGACTGTCCTCCGGCCACCTG AAGTGCCGGCTGAAGATGGACAAGCTGCGGCTGAAGGGCGTGTCCTA CAGCCTGTGTACCGCCGCCTTCACCTTCACCAAGATCCCCGCCGAGAC ACTGCACGGCACCGTGACTGTGGAAGTGCAGTACGCCGGCACCGACG GCCCTTGTAAAGTGCCTGCTCAGATGGCCGTGGATATGCAGACCCTGA CCCCCGTGGGCAGACTGATCACCGCCAACCCTGTGATCACCGAGAGCA CCGAGAACAGCAAGATGATGCTGGAACTGGACCCCCCCTTCGGCGACT CCTACATCGTGATCGGCGTGGGAGAGAAGAAGATCACCCACCACTGGC ACAGATCCGGCAGCACCATCGGCAAGGCCTTTGAGGCTACAGTGCGGG GAGCCAAGAGAATGGCCGTGCTGGGCGATACCGCCTGGGATTTTGGCT CTGTGGGCGGAGCCCTGAACAGCCTGGGAAAGGGCATCCACCAGATCT TCGGCGCTGCCTTCAAGAGCCTGTTCGGCGGCATGAGCTGGTTCAGCC AGATCCTGATCGGCACCCTGCTCGTGTGGCTGGGCCTGAACACCAAGA ACGGCAGCATCTCCCTGACCTGCCTGGCTCTGGGCGGCGTGCTGATCTT TCTGAGCACAGCCGTGTCCGCCTGATAATAGGCTGGAGCCTCGGTGGC CATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGC ACCCGTACCCCCGTGGTCTTTGAATAAAGTCTGAGTGGGCGGCAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAATCTAG IgE HC signal TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGG 122 peptide_prM-E #2 AAATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCAT (Brazil_isolate_Zik GGACTGGACCTGGATCCTGTTCCTGGTGGCCGCTGCCACAAGAGTGCA aSPH2015), CAGCACCAGAAGAGGCAGCGCCTACTACATGTACCTGGACAGAAGCG Sequence, NT (5′ ACGCCGGCGAGGCCATCAGCTTTCCAACCACCCTGGGCATGAACAAGT UTR, ORF, 3′ GCTACATCCAGATCATGGACCTGGGCCACATGTGCGACGCCACCATGA UTR) GCTACGAGTGCCCCATGCTGGACGAGGGCGTGGAACCCGACGATGTG GACTGCTGGTGCAACACCACCAGCACCTGGGTGGTGTACGGCACCTGT CACCACAAGAAGGGCGAAGCCAGACGGTCCAGACGGGCCGTGACACT GCCTAGCCACTCCACCAGAAAGCTGCAGACCCGGTCCCAGACCTGGCT GGAAAGCAGAGAGTACACCAAGCACCTGATCCGGGTGGAAAACTGGA TCTTCCGGAACCCCGGCTTTGCCCTGGCTGCCGCTGCTATTGCTTGGCT GCTGGGCAGCAGCACCTCCCAGAAAGTGATCTACCTCGTGATGATCCT GCTGATCGCCCCTGCCTACAGCATCCGGTGTATCGGCGTGTCCAACCG GGACTTCGTGGAAGGCATGAGCGGCGGCACATGGGTGGACGTGGTGC TGGAACATGGCGGCTGCGTGACAGTGATGGCCCAGGATAAGCCCGCC GTGGACATCGAGCTCGTGACCACCACCGTGTCCAATATGGCCGAAGTG CGGAGCTACTGCTACGAGGCCAGCATCAGCGACATGGCCAGCGACAG CAGATGCCCTACACAGGGCGAGGCCTACCTGGATAAGCAGTCCGACAC CCAGTACGTGTGCAAGCGGACCCTGGTGGATAGAGGCTGGGGCAATG GCTGCGGCCTGTTTGGCAAGGGCAGCCTCGTGACCTGCGCCAAGTTCG CCTGCAGCAAGAAGATGACCGGCAAGAGCATCCAGCCCGAGAACCTG GAATACCGGATCATGCTGAGCGTGCACGGCTCCCAGCACAGCGGCATG ATCGTGAACGACACCGGCCACGAGACAGACGAGAACCGGGCCAAGGT GGAAATCACCCCCAACAGCCCTAGAGCCGAGGCCACACTGGGCGGCTT TGGATCTCTGGGCCTGGACTGCGAGCCTAGAACCGGCCTGGATTTCAG CGACCTGTACTACCTGACCATGAACAACAAGCACTGGCTGGTGCACAA AGAGTGGTTCCACGACATCCCCCTGCCCTGGCATGCTGGCGCTGATAC AGGCACCCCCCACTGGAACAACAAAGAGGCTCTGGTGGAATTCAAGG ACGCCCACGCCAAGCGGCAGACCGTGGTGGTGCTGGGATCTCAGGAA GGCGCCGTGCATACAGCTCTGGCTGGCGCCCTGGAAGCCGAAATGGAT GGCGCCAAAGGCAGACTGTCCTCCGGCCACCTGAAGTGCCGGCTGAAG ATGGACAAGCTGCGGCTGAAGGGCGTGTCCTACAGCCTGTGTACCGCC GCCTTCACCTTCACCAAGATCCCCGCCGAGACACTGCACGGCACCGTG ACTGTGGAAGTGCAGTACGCCGGCACCGACGGCCCTTGTAAAGTGCCT GCTCAGATGGCCGTGGATATGCAGACCCTGACCCCCGTGGGCAGACTG ATCACCGCCAACCCTGTGATCACCGAGAGCACCGAGAACAGCAAGAT GATGCTGGAACTGGACCCCCCCTTCGGCGACTCCTACATCGTGATCGG CGTGGGAGAGAAGAAGATCACCCACCACTGGCACAGATCCGGCAGCA CCATCGGCAAGGCCTTTGAGGCTACAGTGCGGGGAGCCAAGAGAATG GCCGTGCTGGGCGATACCGCCTGGGATTTTGGCTCTGTGGGCGGAGCC CTGAACAGCCTGGGAAAGGGCATCCACCAGATCTTCGGAGCCGCCTTT AAGAGCCTGTTCGGCGGCATGAGCTGGTTCAGCCAGATCCTGATCGGC ACCCTGCTCGTGTGGCTGGGCCTGAACACCAAGAACGGCAGCATCTCC CTGACCTGCCTGGCTCTGGGCGGCGTGCTGATCTTTCTGAGCACAGCC GTGTCCGCCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCC CTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCG TGGTCTTTGAATAAAGTCTGAGTGGGCGGC IgE HC signal ATGGACTGGACCTGGATCCTGTTCCTGGTGGCCGCTGCCACAAGAGTG 123 peptide_prM-E #2 CACAGCACCAGAAGAGGCAGCGCCTACTACATGTACCTGGACAGAAG (Brazil_isolate_ CGACGCCGGCGAGGCCATCAGCTTTCCAACCACCCTGGGCATGAACAA ZikaSPH2015), ORF GTGCTACATCCAGATCATGGACCTGGGCCACATGTGCGACGCCACCAT Sequence, NT GAGCTACGAGTGCCCCATGCTGGACGAGGGCGTGGAACCCGACGATG TGGACTGCTGGTGCAACACCACCAGCACCTGGGTGGTGTACGGCACCT GTCACCACAAGAAGGGCGAAGCCAGACGGTCCAGACGGGCCGTGACA CTGCCTAGCCACTCCACCAGAAAGCTGCAGACCCGGTCCCAGACCTGG CTGGAAAGCAGAGAGTACACCAAGCACCTGATCCGGGTGGAAAACTG GATCTTCCGGAACCCCGGCTTTGCCCTGGCTGCCGCTGCTATTGCTTGG CTGCTGGGCAGCAGCACCTCCCAGAAAGTGATCTACCTCGTGATGATC CTGCTGATCGCCCCTGCCTACAGCATCCGGTGTATCGGCGTGTCCAACC GGGACTTCGTGGAAGGCATGAGCGGCGGCACATGGGTGGACGTGGTG CTGGAACATGGCGGCTGCGTGACAGTGATGGCCCAGGATAAGCCCGCC GTGGACATCGAGCTCGTGACCACCACCGTGTCCAATATGGCCGAAGTG CGGAGCTACTGCTACGAGGCCAGCATCAGCGACATGGCCAGCGACAG CAGATGCCCTACACAGGGCGAGGCCTACCTGGATAAGCAGTCCGACAC CCAGTACGTGTGCAAGCGGACCCTGGTGGATAGAGGCTGGGGCAATG GCTGCGGCCTGTTTGGCAAGGGCAGCCTCGTGACCTGCGCCAAGTTCG CCTGCAGCAAGAAGATGACCGGCAAGAGCATCCAGCCCGAGAACCTG GAATACCGGATCATGCTGAGCGTGCACGGCTCCCAGCACAGCGGCATG ATCGTGAACGACACCGGCCACGAGACAGACGAGAACCGGGCCAAGGT GGAAATCACCCCCAACAGCCCTAGAGCCGAGGCCACACTGGGCGGCTT TGGATCTCTGGGCCTGGACTGCGAGCCTAGAACCGGCCTGGATTTCAG CGACCTGTACTACCTGACCATGAACAACAAGCACTGGCTGGTGCACAA AGAGTGGTTCCACGACATCCCCCTGCCCTGGCATGCTGGCGCTGATAC AGGCACCCCCCACTGGAACAACAAAGAGGCTCTGGTGGAATTCAAGG ACGCCCACGCCAAGCGGCAGACCGTGGTGGTGCTGGGATCTCAGGAA GGCGCCGTGCATACAGCTCTGGCTGGCGCCCTGGAAGCCGAAATGGAT GGCGCCAAAGGCAGACTGTCCTCCGGCCACCTGAAGTGCCGGCTGAAG ATGGACAAGCTGCGGCTGAAGGGCGTGTCCTACAGCCTGTGTACCGCC GCCTTCACCTTCACCAAGATCCCCGCCGAGACACTGCACGGCACCGTG ACTGTGGAAGTGCAGTACGCCGGCACCGACGGCCCTTGTAAAGTGCCT GCTCAGATGGCCGTGGATATGCAGACCCTGACCCCCGTGGGCAGACTG ATCACCGCCAACCCTGTGATCACCGAGAGCACCGAGAACAGCAAGAT GATGCTGGAACTGGACCCCCCCTTCGGCGACTCCTACATCGTGATCGG CGTGGGAGAGAAGAAGATCACCCACCACTGGCACAGATCCGGCAGCA CCATCGGCAAGGCCTTTGAGGCTACAGTGCGGGGAGCCAAGAGAATG GCCGTGCTGGGCGATACCGCCTGGGATTTTGGCTCTGTGGGCGGAGCC CTGAACAGCCTGGGAAAGGGCATCCACCAGATCTTCGGAGCCGCCTTT AAGAGCCTGTTCGGCGGCATGAGCTGGTTCAGCCAGATCCTGATCGGC ACCCTGCTCGTGTGGCTGGGCCTGAACACCAAGAACGGCAGCATCTCC CTGACCTGCCTGGCTCTGGGCGGCGTGCTGATCTTTCTGAGCACAGCC GTGTCCGCC IgE HC signal G*GGGAAATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCC 124 peptide_prM-E #2 ACCATGGACTGGACCTGGATCCTGTTCCTGGTGGCCGCTGCCACAAGA (Brazil_isolate_ GTGCACAGCACCAGAAGAGGCAGCGCCTACTACATGTACCTGGACAG ZikaSPH2015), mRNA AAGCGACGCCGGCGAGGCCATCAGCTTTCCAACCACCCTGGGCATGAA Sequence (T100 CAAGTGCTACATCCAGATCATGGACCTGGGCCACATGTGCGACGCCAC tail) CATGAGCTACGAGTGCCCCATGCTGGACGAGGGCGTGGAACCCGACG ATGTGGACTGCTGGTGCAACACCACCAGCACCTGGGTGGTGTACGGCA CCTGTCACCACAAGAAGGGCGAAGCCAGACGGTCCAGACGGGCCGTG ACACTGCCTAGCCACTCCACCAGAAAGCTGCAGACCCGGTCCCAGACC TGGCTGGAAAGCAGAGAGTACACCAAGCACCTGATCCGGGTGGAAAA CTGGATCTTCCGGAACCCCGGCTTTGCCCTGGCTGCCGCTGCTATTGCT TGGCTGCTGGGCAGCAGCACCTCCCAGAAAGTGATCTACCTCGTGATG ATCCTGCTGATCGCCCCTGCCTACAGCATCCGGTGTATCGGCGTGTCCA ACCGGGACTTCGTGGAAGGCATGAGCGGCGGCACATGGGTGGACGTG GTGCTGGAACATGGCGGCTGCGTGACAGTGATGGCCCAGGATAAGCCC GCCGTGGACATCGAGCTCGTGACCACCACCGTGTCCAATATGGCCGAA GTGCGGAGCTACTGCTACGAGGCCAGCATCAGCGACATGGCCAGCGA CAGCAGATGCCCTACACAGGGCGAGGCCTACCTGGATAAGCAGTCCG ACACCCAGTACGTGTGCAAGCGGACCCTGGTGGATAGAGGCTGGGGC AATGGCTGCGGCCTGTTTGGCAAGGGCAGCCTCGTGACCTGCGCCAAG TTCGCCTGCAGCAAGAAGATGACCGGCAAGAGCATCCAGCCCGAGAA CCTGGAATACCGGATCATGCTGAGCGTGCACGGCTCCCAGCACAGCGG CATGATCGTGAACGACACCGGCCACGAGACAGACGAGAACCGGGCCA AGGTGGAAATCACCCCCAACAGCCCTAGAGCCGAGGCCACACTGGGC GGCTTTGGATCTCTGGGCCTGGACTGCGAGCCTAGAACCGGCCTGGAT TTCAGCGACCTGTACTACCTGACCATGAACAACAAGCACTGGCTGGTG CACAAAGAGTGGTTCCACGACATCCCCCTGCCCTGGCATGCTGGCGCT GATACAGGCACCCCCCACTGGAACAACAAAGAGGCTCTGGTGGAATTC AAGGACGCCCACGCCAAGCGGCAGACCGTGGTGGTGCTGGGATCTCA GGAAGGCGCCGTGCATACAGCTCTGGCTGGCGCCCTGGAAGCCGAAAT GGATGGCGCCAAAGGCAGACTGTCCTCCGGCCACCTGAAGTGCCGGCT GAAGATGGACAAGCTGCGGCTGAAGGGCGTGTCCTACAGCCTGTGTAC CGCCGCCTTCACCTTCACCAAGATCCCCGCCGAGACACTGCACGGCAC CGTGACTGTGGAAGTGCAGTACGCCGGCACCGACGGCCCTTGTAAAGT GCCTGCTCAGATGGCCGTGGATATGCAGACCCTGACCCCCGTGGGCAG ACTGATCACCGCCAACCCTGTGATCACCGAGAGCACCGAGAACAGCA AGATGATGCTGGAACTGGACCCCCCCTTCGGCGACTCCTACATCGTGA TCGGCGTGGGAGAGAAGAAGATCACCCACCACTGGCACAGATCCGGC AGCACCATCGGCAAGGCCTTTGAGGCTACAGTGCGGGGAGCCAAGAG AATGGCCGTGCTGGGCGATACCGCCTGGGATTTTGGCTCTGTGGGCGG AGCCCTGAACAGCCTGGGAAAGGGCATCCACCAGATCTTCGGAGCCGC CTTTAAGAGCCTGTTCGGCGGCATGAGCTGGTTCAGCCAGATCCTGAT CGGCACCCTGCTCGTGTGGCTGGGCCTGAACACCAAGAACGGCAGCAT CTCCCTGACCTGCCTGGCTCTGGGCGGCGTGCTGATCTTTCTGAGCACA GCCGTGTCCGCCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTT GCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACC CCCGTGGTCTTTGAATAAAGTCTGAGTGGGCGGCAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAATCTA G HuIgG_(k) signal TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGG 125 peptide_prME #1 AAATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCAT (Brazil_isolate_ GGAAACCCCTGCCCAGCTGCTGTTCCTGCTGCTGCTGTGGCTGCCTGAT ZikaSPH2015), ACCACCGGCGTGGAAGTGACCAGAAGAGGCAGCGCCTACTACATGTA Sequence, NT (5′ CCTGGACAGAAGCGACGCCGGCGAGGCCATCAGCTTTCCAACCACCCT UTR, ORF, 3′ GGGCATGAACAAGTGCTACATCCAGATCATGGACCTGGGCCACATGTG UTR) CGACGCCACCATGAGCTACGAGTGCCCCATGCTGGACGAGGGCGTGG AACCCGACGATGTGGACTGCTGGTGCAACACCACCAGCACCTGGGTGG TGTACGGCACCTGTCACCACAAGAAGGGCGAAGCCAGACGGTCCAGA CGGGCCGTGACACTGCCTAGCCACTCCACCAGAAAGCTGCAGACCCGG TCCCAGACCTGGCTGGAAAGCAGAGAGTACACCAAGCACCTGATCCG GGTGGAAAACTGGATCTTCCGGAACCCCGGCTTTGCCCTGGCCGCTGC TGCTATTGCTTGGCTGCTGGGCAGCAGCACCTCCCAGAAAGTGATCTA CCTCGTGATGATCCTGCTGATCGCCCCTGCCTACAGCATCCGGTGTATC GGCGTGTCCAACCGGGACTTCGTGGAAGGCATGAGCGGCGGCACATG GGTGGACGTGGTGCTGGAACATGGCGGCTGCGTGACAGTGATGGCCCA GGATAAGCCCGCCGTGGACATCGAGCTCGTGACCACCACCGTGTCCAA TATGGCCGAAGTGCGGAGCTACTGCTACGAGGCCAGCATCAGCGACAT GGCCAGCGACAGCAGATGCCCTACACAGGGCGAGGCCTACCTGGATA AGCAGTCCGACACCCAGTACGTGTGCAAGCGGACCCTGGTGGATAGA GGCTGGGGCAATGGCTGCGGCCTGTTTGGCAAGGGCAGCCTCGTGACC TGCGCCAAGTTCGCCTGCAGCAAGAAGATGACCGGCAAGAGCATCCA GCCCGAGAACCTGGAATACCGGATCATGCTGAGCGTGCACGGCAGCC AGCACTCCGGCATGATCGTGAACGACACCGGCCACGAGACAGACGAG AACCGGGCCAAGGTGGAAATCACCCCCAACAGCCCTAGAGCCGAGGC CACACTGGGCGGCTTTGGATCTCTGGGCCTGGACTGCGAGCCTAGAAC CGGCCTGGATTTCAGCGACCTGTACTACCTGACCATGAACAACAAGCA CTGGCTGGTGCACAAAGAGTGGTTCCACGACATCCCCCTGCCCTGGCA TGCCGGCGCTGATACAGGCACACCCCACTGGAACAACAAAGAGGCTCT GGTGGAATTCAAGGACGCCCACGCCAAGCGGCAGACCGTGGTGGTGC TGGGATCTCAGGAAGGCGCCGTGCATACAGCTCTGGCTGGCGCCCTGG AAGCCGAAATGGATGGCGCCAAAGGCAGACTGTCCAGCGGCCACCTG AAGTGCCGGCTGAAGATGGACAAGCTGCGGCTGAAGGGCGTGTCCTA CAGCCTGTGTACCGCCGCCTTCACCTTCACCAAGATCCCCGCCGAGAC ACTGCACGGCACCGTGACTGTGGAAGTGCAGTACGCCGGCACCGACG GCCCTTGTAAAGTGCCTGCTCAGATGGCCGTGGATATGCAGACCCTGA CCCCCGTGGGCAGACTGATCACCGCCAACCCTGTGATCACCGAGAGCA CCGAGAACAGCAAGATGATGCTGGAACTGGACCCCCCCTTCGGCGACT CCTACATCGTGATCGGCGTGGGAGAGAAGAAGATCACCCACCACTGGC ACCGCAGCGGCAGCACAATCGGCAAGGCCTTTGAAGCCACAGTGCGG GGAGCCAAGAGAATGGCCGTGCTGGGAGATACCGCCTGGGACTTTGG CTCTGTGGGCGGAGCCCTGAACTCTCTGGGCAAGGGAATCCACCAGAT CTTCGGAGCCGCCTTTAAGAGCCTGTTCGGCGGCATGAGCTGGTTCAG CCAGATCCTGATCGGCACCCTGCTCGTGTGGCTGGGCCTGAACACCAA GAACGGCAGCATCTCCCTGACCTGCCTGGCTCTGGGAGGCGTGCTGAT CTTTCTGAGCACCGCCGTGTCTGCCTGATAATAGGCTGGAGCCTCGGT GGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTC CTGCACCCGTACCCCCGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC HuIgG_(k) signal ATGGAAACCCCTGCCCAGCTGCTGTTCCTGCTGCTGCTGTGGCTGCCTG 126 peptide_prME #1 ATACCACCGGCGTGGAAGTGACCAGAAGAGGCAGCGCCTACTACATG (Brazil_isolate_ TACCTGGACAGAAGCGACGCCGGCGAGGCCATCAGCTTTCCAACCACC ZikaSPH2015), ORF CTGGGCATGAACAAGTGCTACATCCAGATCATGGACCTGGGCCACATG Sequence, NT TGCGACGCCACCATGAGCTACGAGTGCCCCATGCTGGACGAGGGCGTG GAACCCGACGATGTGGACTGCTGGTGCAACACCACCAGCACCTGGGTG GTGTACGGCACCTGTCACCACAAGAAGGGCGAAGCCAGACGGTCCAG ACGGGCCGTGACACTGCCTAGCCACTCCACCAGAAAGCTGCAGACCCG GTCCCAGACCTGGCTGGAAAGCAGAGAGTACACCAAGCACCTGATCC GGGTGGAAAACTGGATCTTCCGGAACCCCGGCTTTGCCCTGGCCGCTG CTGCTATTGCTTGGCTGCTGGGCAGCAGCACCTCCCAGAAAGTGATCT ACCTCGTGATGATCCTGCTGATCGCCCCTGCCTACAGCATCCGGTGTAT CGGCGTGTCCAACCGGGACTTCGTGGAAGGCATGAGCGGCGGCACAT GGGTGGACGTGGTGCTGGAACATGGCGGCTGCGTGACAGTGATGGCCC AGGATAAGCCCGCCGTGGACATCGAGCTCGTGACCACCACCGTGTCCA ATATGGCCGAAGTGCGGAGCTACTGCTACGAGGCCAGCATCAGCGAC ATGGCCAGCGACAGCAGATGCCCTACACAGGGCGAGGCCTACCTGGA TAAGCAGTCCGACACCCAGTACGTGTGCAAGCGGACCCTGGTGGATAG AGGCTGGGGCAATGGCTGCGGCCTGTTTGGCAAGGGCAGCCTCGTGAC CTGCGCCAAGTTCGCCTGCAGCAAGAAGATGACCGGCAAGAGCATCC AGCCCGAGAACCTGGAATACCGGATCATGCTGAGCGTGCACGGCAGC CAGCACTCCGGCATGATCGTGAACGACACCGGCCACGAGACAGACGA GAACCGGGCCAAGGTGGAAATCACCCCCAACAGCCCTAGAGCCGAGG CCACACTGGGCGGCTTTGGATCTCTGGGCCTGGACTGCGAGCCTAGAA CCGGCCTGGATTTCAGCGACCTGTACTACCTGACCATGAACAACAAGC ACTGGCTGGTGCACAAAGAGTGGTTCCACGACATCCCCCTGCCCTGGC ATGCCGGCGCTGATACAGGCACACCCCACTGGAACAACAAAGAGGCT CTGGTGGAATTCAAGGACGCCCACGCCAAGCGGCAGACCGTGGTGGT GCTGGGATCTCAGGAAGGCGCCGTGCATACAGCTCTGGCTGGCGCCCT GGAAGCCGAAATGGATGGCGCCAAAGGCAGACTGTCCAGCGGCCACC TGAAGTGCCGGCTGAAGATGGACAAGCTGCGGCTGAAGGGCGTGTCCT ACAGCCTGTGTACCGCCGCCTTCACCTTCACCAAGATCCCCGCCGAGA CACTGCACGGCACCGTGACTGTGGAAGTGCAGTACGCCGGCACCGACG GCCCTTGTAAAGTGCCTGCTCAGATGGCCGTGGATATGCAGACCCTGA CCCCCGTGGGCAGACTGATCACCGCCAACCCTGTGATCACCGAGAGCA CCGAGAACAGCAAGATGATGCTGGAACTGGACCCCCCCTTCGGCGACT CCTACATCGTGATCGGCGTGGGAGAGAAGAAGATCACCCACCACTGGC ACCGCAGCGGCAGCACAATCGGCAAGGCCTTTGAAGCCACAGTGCGG GGAGCCAAGAGAATGGCCGTGCTGGGAGATACCGCCTGGGACTTTGG CTCTGTGGGCGGAGCCCTGAACTCTCTGGGCAAGGGAATCCACCAGAT CTTCGGAGCCGCCTTTAAGAGCCTGTTCGGCGGCATGAGCTGGTTCAG CCAGATCCTGATCGGCACCCTGCTCGTGTGGCTGGGCCTGAACACCAA GAACGGCAGCATCTCCCTGACCTGCCTGGCTCTGGGAGGCGTGCTGAT CTTTCTGAGCACCGCCGTGTCTGCC HuIgG_(k) signal G*GGGAAATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCC 127 peptide_prME #1 ACCATGGAAACCCCTGCCCAGCTGCTGTTCCTGCTGCTGCTGTGGCTGC (Brazil_isolate_ CTGATACCACCGGCGTGGAAGTGACCAGAAGAGGCAGCGCCTACTAC ZikaSPH2015), mRNA ATGTACCTGGACAGAAGCGACGCCGGCGAGGCCATCAGCTTTCCAACC Sequence (T100 ACCCTGGGCATGAACAAGTGCTACATCCAGATCATGGACCTGGGCCAC tail) ATGTGCGACGCCACCATGAGCTACGAGTGCCCCATGCTGGACGAGGGC GTGGAACCCGACGATGTGGACTGCTGGTGCAACACCACCAGCACCTGG GTGGTGTACGGCACCTGTCACCACAAGAAGGGCGAAGCCAGACGGTC CAGACGGGCCGTGACACTGCCTAGCCACTCCACCAGAAAGCTGCAGAC CCGGTCCCAGACCTGGCTGGAAAGCAGAGAGTACACCAAGCACCTGA TCCGGGTGGAAAACTGGATCTTCCGGAACCCCGGCTTTGCCCTGGCCG CTGCTGCTATTGCTTGGCTGCTGGGCAGCAGCACCTCCCAGAAAGTGA TCTACCTCGTGATGATCCTGCTGATCGCCCCTGCCTACAGCATCCGGTG TATCGGCGTGTCCAACCGGGACTTCGTGGAAGGCATGAGCGGCGGCAC ATGGGTGGACGTGGTGCTGGAACATGGCGGCTGCGTGACAGTGATGGC CCAGGATAAGCCCGCCGTGGACATCGAGCTCGTGACCACCACCGTGTC CAATATGGCCGAAGTGCGGAGCTACTGCTACGAGGCCAGCATCAGCG ACATGGCCAGCGACAGCAGATGCCCTACACAGGGCGAGGCCTACCTG GATAAGCAGTCCGACACCCAGTACGTGTGCAAGCGGACCCTGGTGGAT AGAGGCTGGGGCAATGGCTGCGGCCTGTTTGGCAAGGGCAGCCTCGTG ACCTGCGCCAAGTTCGCCTGCAGCAAGAAGATGACCGGCAAGAGCAT CCAGCCCGAGAACCTGGAATACCGGATCATGCTGAGCGTGCACGGCA GCCAGCACTCCGGCATGATCGTGAACGACACCGGCCACGAGACAGAC GAGAACCGGGCCAAGGTGGAAATCACCCCCAACAGCCCTAGAGCCGA GGCCACACTGGGCGGCTTTGGATCTCTGGGCCTGGACTGCGAGCCTAG AACCGGCCTGGATTTCAGCGACCTGTACTACCTGACCATGAACAACAA GCACTGGCTGGTGCACAAAGAGTGGTTCCACGACATCCCCCTGCCCTG GCATGCCGGCGCTGATACAGGCACACCCCACTGGAACAACAAAGAGG CTCTGGTGGAATTCAAGGACGCCCACGCCAAGCGGCAGACCGTGGTGG TGCTGGGATCTCAGGAAGGCGCCGTGCATACAGCTCTGGCTGGCGCCC TGGAAGCCGAAATGGATGGCGCCAAAGGCAGACTGTCCAGCGGCCAC CTGAAGTGCCGGCTGAAGATGGACAAGCTGCGGCTGAAGGGCGTGTC CTACAGCCTGTGTACCGCCGCCTTCACCTTCACCAAGATCCCCGCCGA GACACTGCACGGCACCGTGACTGTGGAAGTGCAGTACGCCGGCACCG ACGGCCCTTGTAAAGTGCCTGCTCAGATGGCCGTGGATATGCAGACCC TGACCCCCGTGGGCAGACTGATCACCGCCAACCCTGTGATCACCGAGA GCACCGAGAACAGCAAGATGATGCTGGAACTGGACCCCCCCTTCGGCG ACTCCTACATCGTGATCGGCGTGGGAGAGAAGAAGATCACCCACCACT GGCACCGCAGCGGCAGCACAATCGGCAAGGCCTTTGAAGCCACAGTG CGGGGAGCCAAGAGAATGGCCGTGCTGGGAGATACCGCCTGGGACTT TGGCTCTGTGGGCGGAGCCCTGAACTCTCTGGGCAAGGGAATCCACCA GATCTTCGGAGCCGCCTTTAAGAGCCTGTTCGGCGGCATGAGCTGGTT CAGCCAGATCCTGATCGGCACCCTGCTCGTGTGGCTGGGCCTGAACAC CAAGAACGGCAGCATCTCCCTGACCTGCCTGGCTCTGGGAGGCGTGCT GATCTTTCTGAGCACCGCCGTGTCTGCCTGATAATAGGCTGGAGCCTC GGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCC TTCCTGCACCCGTACCCCCGTGGTCTTTGAATAAAGTCTGAGTGGGCG GCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAATCTAG HuIgG_(k) signal TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGG 128 peptide_prME #2 AAATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCAT (Brazil_isolate_ GGAAACCCCTGCCCAGCTGCTGTTCCTGCTGCTGCTGTGGCTGCCTGAT ZikaSPH2015), ACCACCGGCACCAGAAGAGGCAGCGCCTACTACATGTACCTGGACAG Sequence, NT (5′ AAGCGACGCCGGCGAGGCCATCAGCTTTCCAACCACCCTGGGCATGAA UTR, ORF, 3′ CAAGTGCTACATCCAGATCATGGACCTGGGCCACATGTGCGACGCCAC UTR) CATGAGCTACGAGTGCCCCATGCTGGACGAGGGCGTGGAACCCGACG ATGTGGACTGCTGGTGCAACACCACCAGCACCTGGGTGGTGTACGGCA CCTGTCACCACAAGAAGGGCGAAGCCAGACGGTCCAGACGGGCCGTG ACACTGCCTAGCCACTCCACCAGAAAGCTGCAGACCCGGTCCCAGACC TGGCTGGAAAGCAGAGAGTACACCAAGCACCTGATCCGGGTGGAAAA CTGGATCTTCCGGAACCCCGGCTTTGCCCTGGCCGCTGCTGCTATTGCT TGGCTGCTGGGCAGCAGCACCTCCCAGAAAGTGATCTACCTCGTGATG ATCCTGCTGATCGCCCCTGCCTACAGCATCCGGTGTATCGGCGTGTCCA ACCGGGACTTCGTGGAAGGCATGAGCGGCGGCACATGGGTGGACGTG GTGCTGGAACATGGCGGCTGCGTGACAGTGATGGCCCAGGATAAGCCC GCCGTGGACATCGAGCTCGTGACCACCACCGTGTCCAATATGGCCGAA GTGCGGAGCTACTGCTACGAGGCCAGCATCAGCGACATGGCCAGCGA CAGCAGATGCCCTACACAGGGCGAGGCCTACCTGGATAAGCAGTCCG ACACCCAGTACGTGTGCAAGCGGACCCTGGTGGATAGAGGCTGGGGC AATGGCTGCGGCCTGTTTGGCAAGGGCAGCCTCGTGACCTGCGCCAAG TTCGCCTGCAGCAAGAAGATGACCGGCAAGAGCATCCAGCCCGAGAA CCTGGAATACCGGATCATGCTGAGCGTGCACGGCAGCCAGCACTCCGG CATGATCGTGAACGACACCGGCCACGAGACAGACGAGAACCGGGCCA AGGTGGAAATCACCCCCAACAGCCCTAGAGCCGAGGCCACACTGGGC GGCTTTGGATCTCTGGGCCTGGACTGCGAGCCTAGAACCGGCCTGGAT TTCAGCGACCTGTACTACCTGACCATGAACAACAAGCACTGGCTGGTG CACAAAGAGTGGTTCCACGACATCCCCCTGCCCTGGCATGCCGGCGCT GATACAGGCACACCCCACTGGAACAACAAAGAGGCTCTGGTGGAATT CAAGGACGCCCACGCCAAGCGGCAGACCGTGGTGGTGCTGGGATCTC AGGAAGGCGCCGTGCATACAGCTCTGGCTGGCGCCCTGGAAGCCGAA ATGGATGGCGCCAAAGGCAGACTGTCCAGCGGCCACCTGAAGTGCCG GCTGAAGATGGACAAGCTGCGGCTGAAGGGCGTGTCCTACAGCCTGTG TACCGCCGCCTTCACCTTCACCAAGATCCCCGCCGAGACACTGCACGG CACCGTGACTGTGGAAGTGCAGTACGCCGGCACCGACGGCCCTTGTAA AGTGCCTGCTCAGATGGCCGTGGATATGCAGACCCTGACCCCCGTGGG CAGACTGATCACCGCCAACCCTGTGATCACCGAGAGCACCGAGAACA GCAAGATGATGCTGGAACTGGACCCCCCCTTCGGCGACTCCTACATCG TGATCGGCGTGGGAGAGAAGAAGATCACCCACCACTGGCACCGCAGC GGCAGCACAATCGGCAAGGCCTTTGAAGCCACAGTGCGGGGAGCCAA GAGAATGGCCGTGCTGGGAGATACCGCCTGGGACTTTGGCTCTGTGGG CGGAGCCCTGAACTCTCTGGGCAAGGGAATCCACCAGATCTTCGGAGC CGCCTTTAAGAGCCTGTTCGGCGGCATGAGCTGGTTCAGCCAGATCCT GATCGGCACCCTGCTCGTGTGGCTGGGCCTGAACACCAAGAACGGCAG CATCTCCCTGACCTGCCTGGCTCTGGGAGGCGTGCTGATCTTTCTGAGC ACCGCCGTGTCTGCCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTT CTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGT ACCCCCGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC HuIgG_(k) signal ATGGAAACCCCTGCCCAGCTGCTGTTCCTGCTGCTGCTGTGGCTGCCTG 129 peptide_prME #2 ATACCACCGGCACCAGAAGAGGCAGCGCCTACTACATGTACCTGGACA (Brazil_isolate_ GAAGCGACGCCGGCGAGGCCATCAGCTTTCCAACCACCCTGGGCATGA ZikaSPH2015), ORF ACAAGTGCTACATCCAGATCATGGACCTGGGCCACATGTGCGACGCCA Sequence, NT CCATGAGCTACGAGTGCCCCATGCTGGACGAGGGCGTGGAACCCGAC GATGTGGACTGCTGGTGCAACACCACCAGCACCTGGGTGGTGTACGGC ACCTGTCACCACAAGAAGGGCGAAGCCAGACGGTCCAGACGGGCCGT GACACTGCCTAGCCACTCCACCAGAAAGCTGCAGACCCGGTCCCAGAC CTGGCTGGAAAGCAGAGAGTACACCAAGCACCTGATCCGGGTGGAAA ACTGGATCTTCCGGAACCCCGGCTTTGCCCTGGCCGCTGCTGCTATTGC TTGGCTGCTGGGCAGCAGCACCTCCCAGAAAGTGATCTACCTCGTGAT GATCCTGCTGATCGCCCCTGCCTACAGCATCCGGTGTATCGGCGTGTCC AACCGGGACTTCGTGGAAGGCATGAGCGGCGGCACATGGGTGGACGT GGTGCTGGAACATGGCGGCTGCGTGACAGTGATGGCCCAGGATAAGC CCGCCGTGGACATCGAGCTCGTGACCACCACCGTGTCCAATATGGCCG AAGTGCGGAGCTACTGCTACGAGGCCAGCATCAGCGACATGGCCAGC GACAGCAGATGCCCTACACAGGGCGAGGCCTACCTGGATAAGCAGTC CGACACCCAGTACGTGTGCAAGCGGACCCTGGTGGATAGAGGCTGGG GCAATGGCTGCGGCCTGTTTGGCAAGGGCAGCCTCGTGACCTGCGCCA AGTTCGCCTGCAGCAAGAAGATGACCGGCAAGAGCATCCAGCCCGAG AACCTGGAATACCGGATCATGCTGAGCGTGCACGGCAGCCAGCACTCC GGCATGATCGTGAACGACACCGGCCACGAGACAGACGAGAACCGGGC CAAGGTGGAAATCACCCCCAACAGCCCTAGAGCCGAGGCCACACTGG GCGGCTTTGGATCTCTGGGCCTGGACTGCGAGCCTAGAACCGGCCTGG ATTTCAGCGACCTGTACTACCTGACCATGAACAACAAGCACTGGCTGG TGCACAAAGAGTGGTTCCACGACATCCCCCTGCCCTGGCATGCCGGCG CTGATACAGGCACACCCCACTGGAACAACAAAGAGGCTCTGGTGGAA TTCAAGGACGCCCACGCCAAGCGGCAGACCGTGGTGGTGCTGGGATCT CAGGAAGGCGCCGTGCATACAGCTCTGGCTGGCGCCCTGGAAGCCGA AATGGATGGCGCCAAAGGCAGACTGTCCAGCGGCCACCTGAAGTGCC GGCTGAAGATGGACAAGCTGCGGCTGAAGGGCGTGTCCTACAGCCTGT GTACCGCCGCCTTCACCTTCACCAAGATCCCCGCCGAGACACTGCACG GCACCGTGACTGTGGAAGTGCAGTACGCCGGCACCGACGGCCCTTGTA AAGTGCCTGCTCAGATGGCCGTGGATATGCAGACCCTGACCCCCGTGG GCAGACTGATCACCGCCAACCCTGTGATCACCGAGAGCACCGAGAAC AGCAAGATGATGCTGGAACTGGACCCCCCCTTCGGCGACTCCTACATC GTGATCGGCGTGGGAGAGAAGAAGATCACCCACCACTGGCACCGCAG CGGCAGCACAATCGGCAAGGCCTTTGAAGCCACAGTGCGGGGAGCCA AGAGAATGGCCGTGCTGGGAGATACCGCCTGGGACTTTGGCTCTGTGG GCGGAGCCCTGAACTCTCTGGGCAAGGGAATCCACCAGATCTTCGGAG CCGCCTTTAAGAGCCTGTTCGGCGGCATGAGCTGGTTCAGCCAGATCC TGATCGGCACCCTGCTCGTGTGGCTGGGCCTGAACACCAAGAACGGCA GCATCTCCCTGACCTGCCTGGCTCTGGGAGGCGTGCTGATCTTTCTGAG CACCGCCGTGTCTGCC HuIgG_(k) signal G*GGGAAATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCC 130 peptide_prME #2 ACCATGGAAACCCCTGCCCAGCTGCTGTTCCTGCTGCTGCTGTGGCTGC (Brazil_isolate_ CTGATACCACCGGCACCAGAAGAGGCAGCGCCTACTACATGTACCTGG ZikaSPH2015), mRNA ACAGAAGCGACGCCGGCGAGGCCATCAGCTTTCCAACCACCCTGGGCA Sequence (T100 TGAACAAGTGCTACATCCAGATCATGGACCTGGGCCACATGTGCGACG tail) CCACCATGAGCTACGAGTGCCCCATGCTGGACGAGGGCGTGGAACCCG ACGATGTGGACTGCTGGTGCAACACCACCAGCACCTGGGTGGTGTACG GCACCTGTCACCACAAGAAGGGCGAAGCCAGACGGTCCAGACGGGCC GTGACACTGCCTAGCCACTCCACCAGAAAGCTGCAGACCCGGTCCCAG ACCTGGCTGGAAAGCAGAGAGTACACCAAGCACCTGATCCGGGTGGA AAACTGGATCTTCCGGAACCCCGGCTTTGCCCTGGCCGCTGCTGCTATT GCTTGGCTGCTGGGCAGCAGCACCTCCCAGAAAGTGATCTACCTCGTG ATGATCCTGCTGATCGCCCCTGCCTACAGCATCCGGTGTATCGGCGTGT CCAACCGGGACTTCGTGGAAGGCATGAGCGGCGGCACATGGGTGGAC GTGGTGCTGGAACATGGCGGCTGCGTGACAGTGATGGCCCAGGATAA GCCCGCCGTGGACATCGAGCTCGTGACCACCACCGTGTCCAATATGGC CGAAGTGCGGAGCTACTGCTACGAGGCCAGCATCAGCGACATGGCCA GCGACAGCAGATGCCCTACACAGGGCGAGGCCTACCTGGATAAGCAG TCCGACACCCAGTACGTGTGCAAGCGGACCCTGGTGGATAGAGGCTGG GGCAATGGCTGCGGCCTGTTTGGCAAGGGCAGCCTCGTGACCTGCGCC AAGTTCGCCTGCAGCAAGAAGATGACCGGCAAGAGCATCCAGCCCGA GAACCTGGAATACCGGATCATGCTGAGCGTGCACGGCAGCCAGCACTC CGGCATGATCGTGAACGACACCGGCCACGAGACAGACGAGAACCGGG CCAAGGTGGAAATCACCCCCAACAGCCCTAGAGCCGAGGCCACACTG GGCGGCTTTGGATCTCTGGGCCTGGACTGCGAGCCTAGAACCGGCCTG GATTTCAGCGACCTGTACTACCTGACCATGAACAACAAGCACTGGCTG GTGCACAAAGAGTGGTTCCACGACATCCCCCTGCCCTGGCATGCCGGC GCTGATACAGGCACACCCCACTGGAACAACAAAGAGGCTCTGGTGGA ATTCAAGGACGCCCACGCCAAGCGGCAGACCGTGGTGGTGCTGGGATC TCAGGAAGGCGCCGTGCATACAGCTCTGGCTGGCGCCCTGGAAGCCGA AATGGATGGCGCCAAAGGCAGACTGTCCAGCGGCCACCTGAAGTGCC GGCTGAAGATGGACAAGCTGCGGCTGAAGGGCGTGTCCTACAGCCTGT GTACCGCCGCCTTCACCTTCACCAAGATCCCCGCCGAGACACTGCACG GCACCGTGACTGTGGAAGTGCAGTACGCCGGCACCGACGGCCCTTGTA AAGTGCCTGCTCAGATGGCCGTGGATATGCAGACCCTGACCCCCGTGG GCAGACTGATCACCGCCAACCCTGTGATCACCGAGAGCACCGAGAAC AGCAAGATGATGCTGGAACTGGACCCCCCCTTCGGCGACTCCTACATC GTGATCGGCGTGGGAGAGAAGAAGATCACCCACCACTGGCACCGCAG CGGCAGCACAATCGGCAAGGCCTTTGAAGCCACAGTGCGGGGAGCCA AGAGAATGGCCGTGCTGGGAGATACCGCCTGGGACTTTGGCTCTGTGG GCGGAGCCCTGAACTCTCTGGGCAAGGGAATCCACCAGATCTTCGGAG CCGCCTTTAAGAGCCTGTTCGGCGGCATGAGCTGGTTCAGCCAGATCC TGATCGGCACCCTGCTCGTGTGGCTGGGCCTGAACACCAAGAACGGCA GCATCTCCCTGACCTGCCTGGCTCTGGGAGGCGTGCTGATCTTTCTGAG CACCGCCGTGTCTGCCTGATAATAGGCTGGAGCCTCGGTGGCCATGCT TCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCG TACCCCCGTGGTCTTTGAATAAAGTCTGAGTGGGCGGCAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAT CTAG HuIgG_(k) signal TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGG 131 peptide_E AAATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCAT (Brazil_isolate_ GGAAACCCCTGCCCAGCTGCTGTTCCTGCTGCTGCTGTGGCTGCCTGAC ZikaSPH2015), ACCACCGGCATCAGATGCATCGGCGTGTCCAACCGGGACTTCGTGGAA Sequence, NT (5′ GGCATGAGCGGCGGCACATGGGTGGACGTGGTGCTGGAACATGGCGG UTR, ORF, 3′ CTGCGTGACAGTGATGGCCCAGGATAAGCCCGCCGTGGACATCGAGCT UTR) CGTGACCACCACCGTGTCCAATATGGCCGAAGTGCGGAGCTACTGCTA CGAGGCCAGCATCAGCGACATGGCCAGCGACAGCAGATGCCCTACAC AGGGCGAGGCCTACCTGGACAAGCAGAGCGACACCCAGTACGTGTGC AAGCGGACCCTGGTGGATAGAGGCTGGGGCAATGGCTGCGGCCTGTTT GGCAAGGGCAGCCTCGTGACCTGCGCCAAGTTCGCCTGCAGCAAGAA GATGACCGGCAAGAGCATCCAGCCCGAGAACCTGGAATACCGGATCA TGCTGAGCGTGCACGGCAGCCAGCACTCCGGCATGATCGTGAACGACA CCGGCCACGAGACAGACGAGAACCGGGCCAAGGTGGAAATCACCCCC AACAGCCCTAGAGCCGAGGCCACACTGGGCGGCTTTGGATCTCTGGGC CTGGACTGCGAGCCTAGAACCGGCCTGGATTTCAGCGACCTGTACTAC CTGACCATGAACAACAAGCACTGGCTGGTGCACAAAGAGTGGTTCCAC GACATCCCCCTGCCCTGGCATGCCGGCGCTGATACAGGCACACCCCAC TGGAACAACAAAGAGGCTCTGGTGGAATTCAAGGACGCCCACGCCAA GCGGCAGACCGTGGTGGTGCTGGGATCTCAGGAAGGCGCCGTGCATAC AGCTCTGGCTGGCGCCCTGGAAGCCGAAATGGATGGCGCCAAAGGCA GACTGAGCAGCGGCCACCTGAAGTGCCGGCTGAAGATGGACAAGCTG CGGCTGAAGGGCGTGTCCTACAGCCTGTGTACCGCCGCCTTCACCTTC ACCAAGATCCCCGCCGAGACACTGCACGGCACCGTGACTGTGGAAGTG CAGTACGCCGGCACCGACGGCCCTTGTAAAGTGCCTGCTCAGATGGCC GTGGATATGCAGACCCTGACCCCCGTGGGCAGGCTGATCACAGCCAAC CCTGTGATCACCGAGAGCACCGAGAACAGCAAGATGATGCTGGAACT GGACCCCCCCTTCGGCGACTCCTACATCGTGATCGGCGTGGGAGAGAA GAAGATCACCCACCACTGGCACAGAAGCGGCAGCACCATCGGCAAGG CCTTTGAGGCTACAGTGCGGGGAGCCAAGAGAATGGCCGTGCTGGGA GATACCGCCTGGGACTTTGGCTCTGTGGGCGGAGCCCTGAACTCTCTG GGCAAGGGAATCCACCAGATCTTCGGCGCTGCCTTCAAGAGCCTGTTC GGCGGCATGAGCTGGTTCAGCCAGATCCTGATCGGCACCCTGCTCGTG TGGCTGGGCCTGAACACCAAGAACGGCAGCATCTCCCTGACCTGCCTG GCTCTGGGAGGCGTGCTGATCTTTCTGAGCACCGCCGTGTCTGCCTGAT AATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCC CCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCGTGGTCTTTGAATA AAGTCTGAGTGGGCGGC HuIgG_(k) signal ATGGAAACCCCTGCCCAGCTGCTGTTCCTGCTGCTGCTGTGGCTGCCTG 132 peptide_E ACACCACCGGCATCAGATGCATCGGCGTGTCCAACCGGGACTTCGTGG (Brazil_isolate_ AAGGCATGAGCGGCGGCACATGGGTGGACGTGGTGCTGGAACATGGC ZikaSPH2015), ORF GGCTGCGTGACAGTGATGGCCCAGGATAAGCCCGCCGTGGACATCGA Sequence, NT GCTCGTGACCACCACCGTGTCCAATATGGCCGAAGTGCGGAGCTACTG CTACGAGGCCAGCATCAGCGACATGGCCAGCGACAGCAGATGCCCTA CACAGGGCGAGGCCTACCTGGACAAGCAGAGCGACACCCAGTACGTG TGCAAGCGGACCCTGGTGGATAGAGGCTGGGGCAATGGCTGCGGCCT GTTTGGCAAGGGCAGCCTCGTGACCTGCGCCAAGTTCGCCTGCAGCAA GAAGATGACCGGCAAGAGCATCCAGCCCGAGAACCTGGAATACCGGA TCATGCTGAGCGTGCACGGCAGCCAGCACTCCGGCATGATCGTGAACG ACACCGGCCACGAGACAGACGAGAACCGGGCCAAGGTGGAAATCACC CCCAACAGCCCTAGAGCCGAGGCCACACTGGGCGGCTTTGGATCTCTG GGCCTGGACTGCGAGCCTAGAACCGGCCTGGATTTCAGCGACCTGTAC TACCTGACCATGAACAACAAGCACTGGCTGGTGCACAAAGAGTGGTTC CACGACATCCCCCTGCCCTGGCATGCCGGCGCTGATACAGGCACACCC CACTGGAACAACAAAGAGGCTCTGGTGGAATTCAAGGACGCCCACGC CAAGCGGCAGACCGTGGTGGTGCTGGGATCTCAGGAAGGCGCCGTGC ATACAGCTCTGGCTGGCGCCCTGGAAGCCGAAATGGATGGCGCCAAA GGCAGACTGAGCAGCGGCCACCTGAAGTGCCGGCTGAAGATGGACAA GCTGCGGCTGAAGGGCGTGTCCTACAGCCTGTGTACCGCCGCCTTCAC CTTCACCAAGATCCCCGCCGAGACACTGCACGGCACCGTGACTGTGGA AGTGCAGTACGCCGGCACCGACGGCCCTTGTAAAGTGCCTGCTCAGAT GGCCGTGGATATGCAGACCCTGACCCCCGTGGGCAGGCTGATCACAGC CAACCCTGTGATCACCGAGAGCACCGAGAACAGCAAGATGATGCTGG AACTGGACCCCCCCTTCGGCGACTCCTACATCGTGATCGGCGTGGGAG AGAAGAAGATCACCCACCACTGGCACAGAAGCGGCAGCACCATCGGC AAGGCCTTTGAGGCTACAGTGCGGGGAGCCAAGAGAATGGCCGTGCT GGGAGATACCGCCTGGGACTTTGGCTCTGTGGGCGGAGCCCTGAACTC TCTGGGCAAGGGAATCCACCAGATCTTCGGCGCTGCCTTCAAGAGCCT GTTCGGCGGCATGAGCTGGTTCAGCCAGATCCTGATCGGCACCCTGCT CGTGTGGCTGGGCCTGAACACCAAGAACGGCAGCATCTCCCTGACCTG CCTGGCTCTGGGAGGCGTGCTGATCTTTCTGAGCACCGCCGTGTCTGCC HuIgG_(k) signal G*GGGAAATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCC 133 peptide_E ACCATGGAAACCCCTGCCCAGCTGCTGTTCCTGCTGCTGCTGTGGCTGC (Brazil_isolate_ CTGACACCACCGGCATCAGATGCATCGGCGTGTCCAACCGGGACTTCG ZikaSPH2015), mRNA TGGAAGGCATGAGCGGCGGCACATGGGTGGACGTGGTGCTGGAACAT Sequence (T100 GGCGGCTGCGTGACAGTGATGGCCCAGGATAAGCCCGCCGTGGACATC tail) GAGCTCGTGACCACCACCGTGTCCAATATGGCCGAAGTGCGGAGCTAC TGCTACGAGGCCAGCATCAGCGACATGGCCAGCGACAGCAGATGCCCT ACACAGGGCGAGGCCTACCTGGACAAGCAGAGCGACACCCAGTACGT GTGCAAGCGGACCCTGGTGGATAGAGGCTGGGGCAATGGCTGCGGCC TGTTTGGCAAGGGCAGCCTCGTGACCTGCGCCAAGTTCGCCTGCAGCA AGAAGATGACCGGCAAGAGCATCCAGCCCGAGAACCTGGAATACCGG ATCATGCTGAGCGTGCACGGCAGCCAGCACTCCGGCATGATCGTGAAC GACACCGGCCACGAGACAGACGAGAACCGGGCCAAGGTGGAAATCAC CCCCAACAGCCCTAGAGCCGAGGCCACACTGGGCGGCTTTGGATCTCT GGGCCTGGACTGCGAGCCTAGAACCGGCCTGGATTTCAGCGACCTGTA CTACCTGACCATGAACAACAAGCACTGGCTGGTGCACAAAGAGTGGTT CCACGACATCCCCCTGCCCTGGCATGCCGGCGCTGATACAGGCACACC CCACTGGAACAACAAAGAGGCTCTGGTGGAATTCAAGGACGCCCACG CCAAGCGGCAGACCGTGGTGGTGCTGGGATCTCAGGAAGGCGCCGTG CATACAGCTCTGGCTGGCGCCCTGGAAGCCGAAATGGATGGCGCCAAA GGCAGACTGAGCAGCGGCCACCTGAAGTGCCGGCTGAAGATGGACAA GCTGCGGCTGAAGGGCGTGTCCTACAGCCTGTGTACCGCCGCCTTCAC CTTCACCAAGATCCCCGCCGAGACACTGCACGGCACCGTGACTGTGGA AGTGCAGTACGCCGGCACCGACGGCCCTTGTAAAGTGCCTGCTCAGAT GGCCGTGGATATGCAGACCCTGACCCCCGTGGGCAGGCTGATCACAGC CAACCCTGTGATCACCGAGAGCACCGAGAACAGCAAGATGATGCTGG AACTGGACCCCCCCTTCGGCGACTCCTACATCGTGATCGGCGTGGGAG AGAAGAAGATCACCCACCACTGGCACAGAAGCGGCAGCACCATCGGC AAGGCCTTTGAGGCTACAGTGCGGGGAGCCAAGAGAATGGCCGTGCT GGGAGATACCGCCTGGGACTTTGGCTCTGTGGGCGGAGCCCTGAACTC TCTGGGCAAGGGAATCCACCAGATCTTCGGCGCTGCCTTCAAGAGCCT GTTCGGCGGCATGAGCTGGTTCAGCCAGATCCTGATCGGCACCCTGCT CGTGTGGCTGGGCCTGAACACCAAGAACGGCAGCATCTCCCTGACCTG CCTGGCTCTGGGAGGCGTGCTGATCTTTCTGAGCACCGCCGTGTCTGCC TGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCT CCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCGTGGTCTTTG AATAAAGTCTGAGTGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAATCTAG Zika_RIO- ATGCTGGGCAGCAACAGCGGCCAGAGAGTGGTGTTCACCATCCTGCTG 134 U1_JEVsp CTGCTGGTGGCCCCTGCCTACAGCGCCGAAGTGACAAGAAGAGGCAG Zika PRME Strain CGCCTACTACATGTACCTGGACCGGAACGATGCCGGCGAGGCCATCAG ascension id: CTTTCCAACCACCCTGGGCATGAACAAGTGCTACATCCAGATCATGGA ANG09399 with CCTGGGCCACATGTGCGACGCCACCATGAGCTACGAGTGCCCCATGCT JEV PRM signal GGACGAGGGCGTGGAACCCGACGATGTGGACTGCTGGTGCAATACCA sequence CCAGCACCTGGGTGGTGTACGGCACCTGTCACCACAAGAAGGGCGAA (optimized) GCCAGACGGTCCAGACGGGCCGTGACACTGCCTAGCCACAGCACCAG AAAGCTGCAGACCCGGTCCCAGACCTGGCTGGAAAGCAGAGAGTACA CCAAGCACCTGATCCGGGTGGAAAACTGGATCTTCCGGAACCCCGGCT TTGCCCTGGCTGCCGCTGCTATTGCTTGGCTGCTGGGCTCTAGCACCAG CCAGAAAGTGATCTACCTCGTGATGATCCTGCTGATCGCCCCAGCCTA CTCCATCCGGTGTATCGGCGTGTCCAACCGGGACTTCGTGGAAGGCAT GAGCGGCGGCACATGGGTGGACGTGGTGCTGGAACATGGCGGCTGCG TGACAGTGATGGCCCAGGACAAGCCCACCGTGGACATCGAGCTCGTGA CCACCACCGTGTCCAATATGGCCGAAGTGCGGAGCTACTGCTACGAGG CCAGCATCAGCGACATGGCCAGCGACAGCAGATGCCCTACACAGGGC GAGGCCTACCTGGACAAGCAGTCCGACACCCAGTACGTGTGCAAGCG GACCCTGGTGGACAGGGGCTGGGGCAATGGCTGTGGCCTGTTTGGCAA GGGCAGCCTCGTGACCTGCGCCAAGTTCGCCTGCAGCAAGAAGATGAC CGGCAAGAGCATCCAGCCCGAGAACCTGGAATACCGGATCATGCTGA GCGTGCACGGCTCCCAGCACAGCGGCATGATCGTGAACGACACCGGCC ACGAGACAGACGAGAACCGGGCCAAGGTGGAAATCACCCCCAACAGC CCTAGAGCCGAGGCCACACTGGGCGGCTTTGGATCTCTGGGCCTGGAC TGCGAGCCTAGAACCGGCCTGGATTTCAGCGACCTGTACTACCTGACC ATGAACAACAAACACTGGCTGGTGCACAAAGAGTGGTTCCACGACATC CCCCTGCCCTGGCATGCTGGCGCTGATACAGGCACCCCCCACTGGAAC AACAAAGAGGCCCTGGTGGAATTCAAGGACGCCCACGCCAAGCGGCA GACCGTGGTGGTGCTGGGATCTCAGGAAGGCGCCGTGCATACAGCTCT GGCTGGCGCCCTGGAAGCCGAAATGGATGGCGCCAAAGGCAGACTGA GCAGCGGCCACCTGAAGTGCCGGCTGAAGATGGACAAGCTGCGGCTG AAGGGCGTGTCCTACAGCCTGTGTACCGCCGCCTTCACCTTCACCAAG ATCCCCGCCGAGACACTGCACGGCACCGTGACTGTGGAAGTGCAGTAC GCCGGCACCGACGGCCCTTGTAAAGTGCCTGCTCAGATGGCCGTGGAT ATGCAGACCCTGACCCCCGTGGGCAGGCTGATCACAGCCAACCCTGTG ATCACCGAGAGCACCGAGAACAGCAAGATGATGCTGGAACTGGACCC CCCCTTCGGCGACTCCTACATCGTGATCGGCGTGGGAGAGAAGAAGAT CACCCACCACTGGCACAGAAGCGGCAGCACCATCGGCAAGGCCTTTGA GGCTACAGTGCGGGGAGCCAAGAGAATGGCCGTGCTGGGCGATACCG CCTGGGATTTTGGCTCTGTGGGCGGAGCCCTGAACAGCCTGGGAAAGG GCATCCACCAGATCTTCGGAGCCGCCTTTAAGAGCCTGTTCGGCGGCA TGAGCTGGTTCAGCCAGATCCTGATCGGCACCCTGCTGATGTGGCTGG GCCTGAACACCAAGAACGGCAGCATCTCCCTGATGTGCCTGGCTCTGG GCGGCGTGCTGATCTTTCTGAGCACAGCCGTGTCCGCC Zika_RIO- ATGAAGTGCCTGCTGTACCTGGCCTTCCTGTTCATCGGCGTGAACTGCG 135 U1_VSVgSp CCGAAGTGACCAGAAGAGGCAGCGCCTACTACATGTACCTGGACCGG Zika PRME Strain AACGATGCCGGCGAGGCCATCAGCTTTCCAACCACCCTGGGCATGAAC ascension id: AAGTGCTACATCCAGATCATGGACCTGGGCCACATGTGCGACGCCACC ANG09399 with ATGAGCTACGAGTGCCCCATGCTGGACGAGGGCGTGGAACCCGACGA VSVg protein TGTGGACTGCTGGTGCAACACCACCAGCACCTGGGTGGTGTACGGCAC signal sequence CTGTCACCACAAGAAGGGCGAAGCCAGACGGTCCAGACGGGCCGTGA (optimized) CACTGCCTAGCCACAGCACCAGAAAGCTGCAGACCCGGTCCCAGACCT GGCTGGAAAGCAGAGAGTACACCAAGCACCTGATCCGGGTGGAAAAC TGGATCTTCCGGAACCCCGGCTTTGCCCTGGCCGCTGCTGCTATTGCTT GGCTGCTGGGCAGCAGCACCTCCCAGAAAGTGATCTACCTCGTGATGA TCCTGCTGATCGCCCCTGCCTACAGCATCCGGTGTATCGGCGTGTCCAA CCGGGACTTCGTGGAAGGCATGAGCGGCGGCACATGGGTGGACGTGG TGCTGGAACATGGCGGCTGCGTGACAGTGATGGCCCAGGACAAGCCC ACCGTGGACATCGAGCTCGTGACCACCACCGTGTCCAATATGGCCGAA GTGCGGAGCTACTGCTACGAGGCCAGCATCAGCGACATGGCCAGCGA CAGCAGATGCCCTACACAGGGCGAGGCCTACCTGGACAAGCAGTCCG ACACCCAGTACGTGTGCAAGCGGACCCTGGTGGATAGAGGCTGGGGC AATGGCTGCGGCCTGTTTGGCAAGGGCAGCCTCGTGACCTGTGCCAAG TTCGCCTGCAGCAAGAAGATGACCGGCAAGAGCATCCAGCCCGAGAA CCTGGAATACCGGATCATGCTGAGCGTGCACGGCAGCCAGCACTCCGG CATGATCGTGAACGACACCGGCCACGAGACAGACGAGAACCGGGCCA AGGTGGAAATCACCCCCAACAGCCCTAGAGCCGAGGCCACACTGGGC GGCTTTGGATCTCTGGGCCTGGACTGCGAGCCTAGAACCGGCCTGGAT TTCAGCGACCTGTACTACCTGACCATGAACAACAAGCACTGGCTGGTG CACAAAGAGTGGTTCCACGACATCCCCCTGCCCTGGCATGCCGGCGCT GATACAGGCACACCCCACTGGAACAACAAAGAGGCTCTGGTGGAATT CAAGGACGCCCACGCCAAGCGGCAGACCGTGGTGGTGCTGGGATCTC AGGAAGGCGCCGTGCATACAGCTCTGGCTGGCGCCCTGGAAGCCGAA ATGGATGGCGCCAAAGGCAGACTGTCCAGCGGCCACCTGAAGTGCAG ACTGAAGATGGACAAGCTGCGGCTGAAGGGCGTGTCCTACAGCCTGTG TACCGCCGCCTTCACCTTCACCAAGATCCCCGCCGAGACACTGCACGG CACCGTGACTGTGGAAGTGCAGTACGCCGGCACCGACGGCCCTTGTAA AGTGCCAGCTCAGATGGCCGTGGATATGCAGACCCTGACCCCCGTGGG CAGACTGATCACCGCCAACCCTGTGATCACCGAGAGCACCGAGAACA GCAAGATGATGCTGGAACTGGACCCCCCCTTCGGCGACTCCTACATCG TGATCGGCGTGGGAGAGAAGAAGATCACCCACCACTGGCACAGAAGC GGCAGCACCATCGGCAAGGCCTTTGAGGCTACAGTGCGGGGAGCCAA GAGAATGGCCGTGCTGGGAGATACCGCCTGGGACTTTGGCTCTGTGGG CGGAGCCCTGAACTCTCTGGGCAAGGGAATCCACCAGATCTTCGGAGC CGCCTTTAAGAGCCTGTTCGGCGGCATGAGCTGGTTCAGCCAGATCCT GATCGGCACCCTGCTGATGTGGCTGGGCCTGAACACCAAGAACGGCAG CATCTCCCTGATGTGCCTGGCTCTGGGAGGCGTGCTGATCTTCCTGAGC ACAGCCGTGTCTGCC ZIKA_PRME_DSP_ ATGGACTGGACCTGGATCCTGTTCCTGGTGGCCGCTGCCACAAGAGTG 136 N154A CACAGCGTGGAAGTGACCAGACGGGGCAGCGCCTACTACATGTACCTG Zika PRME Strain GACAGAAGCGACGCCGGCGAGGCCATCAGCTTTCCAACCACCCTGGGC ascension id: ATGAACAAGTGCTACATCCAGATCATGGACCTGGGCCACATGTGCGAC ACD75819 with GCCACCATGAGCTACGAGTGCCCCATGCTGGACGAGGGCGTGGAACCC IgE signal peptide GACGATGTGGACTGCTGGTGCAACACCACCAGCACCTGGGTGGTGTAC (optimized) GGCACCTGTCACCACAAGAAGGGCGAAGCCAGACGGTCCAGACGGGC CGTGACACTGCCTAGCCACAGCACCAGAAAGCTGCAGACCCGGTCCCA GACCTGGCTGGAAAGCAGAGAGTACACCAAGCACCTGATCCGGGTGG AAAACTGGATCTTCCGGAACCCCGGCTTTGCCCTGGCTGCCGCTGCTAT TGCTTGGCTGCTGGGCAGCAGCACCTCCCAGAAAGTGATCTACCTCGT GATGATCCTGCTGATCGCCCCTGCCTACAGCATCCGGTGTATCGGCGT GTCCAACCGGGACTTCGTGGAAGGCATGAGCGGCGGCACATGGGTGG ACGTGGTGCTGGAACATGGCGGCTGCGTGACAGTGATGGCCCAGGATA AGCCCGCCGTGGACATCGAGCTCGTGACCACCACCGTGTCCAATATGG CCGAAGTGCGGAGCTACTGCTACGAGGCCAGCATCAGCGACATGGCC AGCGACAGCAGATGCCCTACACAGGGCGAGGCCTACCTGGATAAGCA GTCCGACACCCAGTACGTGTGCAAGCGGACCCTGGTGGATAGAGGCTG GGGCAATGGCTGCGGCCTGTTTGGCAAGGGCAGCCTCGTGACCTGCGC CAAGTTCGCCTGCAGCAAGAAGATGACCGGCAAGAGCATCCAGCCCG AGAACCTGGAATACCGGATCATGCTGAGCGTGCACGGCTCCCAGCACA GCGGCATGATCGTGGCCGACACCGGCCACGAGACAGACGAGAACCGG GCCAAGGTGGAAATCACCCCCAACAGCCCTAGAGCCGAGGCCACACT GGGCGGCTTTGGATCTCTGGGCCTGGACTGCGAGCCTAGAACCGGCCT GGATTTCAGCGACCTGTACTACCTGACCATGAACAACAAGCACTGGCT GGTGCACAAAGAGTGGTTCCACGACATCCCCCTGCCCTGGCATGCTGG CGCTGATACAGGCACCCCCCACTGGAACAACAAAGAGGCTCTGGTGG AATTCAAGGACGCCCACGCCAAGCGGCAGACCGTGGTGGTGCTGGGA TCTCAGGAAGGCGCCGTGCATACAGCTCTGGCTGGCGCCCTGGAAGCC GAAATGGATGGCGCCAAAGGCAGACTGTCCTCCGGCCACCTGAAGTGC CGGCTGAAGATGGACAAGCTGCGGCTGAAGGGCGTGTCCTACAGCCTG TGTACCGCCGCCTTCACCTTCACCAAGATCCCCGCCGAGACACTGCAC GGCACCGTGACTGTGGAAGTGCAGTACGCCGGCACCGACGGCCCTTGT AAAGTGCCTGCTCAGATGGCCGTGGATATGCAGACCCTGACCCCCGTG GGCAGACTGATCACCGCCAACCCTGTGATCACCGAGAGCACCGAGAA CAGCAAGATGATGCTGGAACTGGACCCCCCCTTCGGCGACTCCTACAT CGTGATCGGCGTGGGAGAGAAGAAGATCACCCACCACTGGCACAGAT CCGGCAGCACCATCGGCAAGGCCTTTGAGGCTACAGTGCGGGGAGCC AAGAGAATGGCCGTGCTGGGCGATACCGCCTGGGATTTTGGCTCTGTG GGCGGAGCCCTGAACAGCCTGGGAAAGGGCATCCACCAGATCTTCGG CGCTGCCTTCAAGAGCCTGTTCGGCGGCATGAGCTGGTTCAGCCAGAT CCTGATCGGCACCCTGCTCGTGTGGCTGGGCCTGAACACCAAGAACGG CAGCATCTCCCTGACCTGCCTGGCTCTGGGCGGCGTGCTGATCTTTCTG AGCACAGCCGTGTCCGCC ZIKV mRNA Sequences Zika virus strain AUGAAAAACCCAAAGAAGAAAUCCGGAGGAUUCCGGAUUGUCAAUA 137 MR 766 UGCUAAAACGCGGAGUAGCCCGUGUAAACCCCUUGGGAGGUUUGAA polyprotein gene, GAGGCUGCCAGCCGGACUUCUGCUGGGUCAUGGACCCAUCAGAAUG complete cds GUUUUGGCGAUAUUAGCCUUUUUGAGAUUCACAGCAAUCAAGCCAU GenBank CACUGGGCCUCAUCAACAGAUGGGGUACCGUGGGGAAAAAAGAGGC Accession: UAUGGAAAUAAUAAAAAAAUUUAAGAAAGAUCUUGCUGCCAUGUUG DQ859059 AGAAUAAUCAAUGCUAGGAAGGAGAGGAAGAGACGUGGCGCAGACA CCAGCAUCGGAAUCGUUGGCCUCCUGUUGACUACAGCCAUGGCAGCA GAGAUCACUAGACGUGGGAGUGCAUACUACAUGUACUUGGAUAGGA GCGAUGCAGGGAAGGCCAUUUCUUUCGCUACCACAUUGGGGGUGAA CAAAUGCCAUGUGCAGAUCAUGGACCUCGGGCACAUGUGUGACGCC ACCAUGAGCUAUGAAUGCCCUAUGCUGGACGAGGGGGUGGAACCAG AUGACGUCGAUUGCUGGUGCAACACGACAUCAACUUGGGUUGUGUA CGGAACCUGUCAUCAUAAAAAAGGUGAAGCACGGCGAUCUAGAAGA GCCGUCACGCUCCCAUCUCACUCCACAAGGAAAUUGCAAACGCGGUC GCAGACUUGGCUAGAAUCAAGAGAAUACACAAAGCACCUGAUCAAG GUUGAAAAUUGGAUAUUCAGGAACCCUGGUUUUACGCUAGUGGCUG UCGCCAUCGCCUGGCUUUUGGGAAGCUCGACGAGCCAAAAAGUCAU AUACUUGGUCAUGAUACUGCUGAUUGCCCCGGCAUACAGUAUCAGG UGCAUAGGAGUCAGCAAUAGAGACUUCGUGGAGGGCAUGUCAGGUG GGACCUGGGUUGACGUUGUCCUGGAACAUGGAGGCUGCGUCACCGU GAUGGCACAGGACAAGCCAACAGUUGACAUAGAGCUGGUCACAACA ACGGUUAGUAACAUGGCCGAGGUGAGAUCCUAUUGUUACGAGGCAU CAAUAUCGGACAUGGCUUCGGACAGUCGCUGCCCAACACAAGGUGA AGCCUACCUUGACAAGCAAUCAGACACUCAAUAUGUUUGCAAAAGA ACAUUGGUGGACAGAGGUUGGGGAAAUGGGUGUGGACUCUUUGGCA AAGGGAGUUUGGUGACAUGUGCUAAGUUCACGUGCUCCAAGAAGAU GACUGGGAAGAGCAUUCAGCCGGAGAACCUGGAGUAUCGGAUAAUG CUAUCAGUGCAUGGCUCCCAGCACAGUGGGAUGAUUGUUAAUGAUG AAAACAGAGCGAAGGUCGAGGUUACGCCCAAUUCACCAAGAGCAGA AGCAACCCUGGGAGGCUUUGGAAGCUUAGGACUUGAUUGUGAACCA AGGACAGGCCUUGACUUUUCAGAUCUGUAUUACCUAACCAUGAAUA ACAAGCAUUGGUUGGUGCACAAAGAGUGGUUUCAUGACAUCCCAUU GCCCUGGCAUGCUGGGGCAGACACUGGAACUCCACAUUGGAACAAC AAGGAGGCAUUAGUGGAAUUCAAGGACGCCCACGCCAAGAGGCAAA CCGUCGUGGUUUUGGGGAGCCAGGAAGGAGCCGUCCACACGGCUCU UGCUGGAGCUCUAGAGGCUGAGAUGGAUGGUGCAAAGGGAAGGCUA UUCUCUGGCCACUUGAAAUGUCGCUUAAAAAUGGACAAGCUUAGAU UGAAGGGCGUGUCAUAUUCCUUGUGCACCGCGGCAUUCACAUUCAC CAAGGUCCCGGCUGAAACACUACAUGGAACAGUCACAGUGGAGGUG CAGUAUGCAGGGACAGAUGGACCCUGCAAGGUCCCAGCCCAGAUGG CGGUGGACAUGCAGACCUUGACCCCAGUCGGAAGGCUGAUAACCGCC AACCCCGUGAUUACUGAAAGCACUGAGAAUUCAAAGAUGAUGUUGG AGCUCGACCCACCAUUUGGGGAUUCUUACAUUGUCAUAGGAGUUGG GGAUAAGAAAAUCACCCAUCACUGGCAUAGGAGUGGCAGCACCAUU GGAAAAGCAUUUGAAGCCACUGUGAGAGGCGCUAAGAGAAUGGCAG UCCUGGGGGACACAGCUUGGGACUUUGGAUCAGUCGGAGGUGUGUU UAACUCAUUGGGCAAGGGCAUUCAUCAGAUUUUUGGAGCAGCUUUC AAAUCACUGUUUGGAGGAAUGUCCUGGUUCUCACAGAUCCUCAUAG GCACUCUGCUGGUGUGGUUAGGUCUGAACACAAAGAAUGGGUCUAU CUCCCUCACAUGCUUAGCCCUGGGGGGAGUGAUGAUCUUCCUCUCCA CGGCUGUUUCUGCUGACGUGGGGUGCUCGGUGGACUUCUCAAAAAA AGAAACGAGAUGUGGCACGGGGGUGUUCGUCUACAAUGACGUUGAA GCCUGGAGGGACCGGUACAAGUACCAUCCUGACUCCCCUCGUAGACU GGCAGCAGCCGUUAAGCAAGCUUGGGAAGAGGGGAUUUGUGGGAUC UCCUCUGUUUCUAGAAUGGAAAACAUAAUGUGGAAAUCAGUGGAAG GAGAGCUCAAUGCAAUCCUAGAGGAGAAUGGAGUCCAACUGACAGU UGUUGUGGGAUCUGUAAAAAACCCCAUGUGGAGAGGCCCACAAAGA UUGCCAGUGCCUGUGAAUGAGCUGCCCCAUGGCUGGAAAGCCUGGG GGAAAUCGUACUUUGUUAGGGCGGCAAAGACCAACAACAGUUUUGU UGUCGACGGUGACACAUUGAAGGAAUGUCCGCUCAAGCACAGAGCA UGGAACAGCUUCCUCGUGGAGGAUCACGGGUUUGGGGUCUUCCACA CCAGUGUUUGGCUUAAGGUUAGAGAAGAUUACUCACUGGAGUGUGA CCCAGCCGUCAUAGGAACAGCUGUUAAGGGAAAGGAGGCCGCGCAC AGUGAUCUAGGCUAUUGGAUUGAAAGUGAAAAGAAUGACACAUGGA GGCUGAAGAGGGCUCAUUUGAUUGAGAUGAAAACAUGUGAGUGGCC AAAGUCUCACACACUGUGGACAGAUGGAGUGGAAGAAAGUGAUCUG AUCAUACCCAAGUCUUUAGCUGGUCCACUCAGCCACCACAACACCAG AGAGGGUUACAGAACUCAAGUGAAAGGGCCAUGGCAUAGUGAGGAG CUUGAAAUCCGAUUUGAGGAAUGUCCAGGUACCAAGGUUCAUGUGG AGGAGACAUGCGGAACGAGAGGACCAUCUCUGAGAUCAACCACUGC AAGCGGAAGGGUCAUUGAGGAAUGGUGCUGUAGGGAAUGCACAAUG CCCCCACUAUCGUUCCGAGCAAAAGAUGGCUGCUGGUAUGGAAUGG AGAUAAGGCCUAGGAAAGAACCAGAGAGCAACUUAGUGAGGUCAAU GGUGACAGCGGGAUCAACCGAUCAUAUGGAUCAUUUUUCUCUUGGA GUGCUUGUGAUUCUACUCAUGGUGCAGGAAGGGUUGAAGAAGAGAA UGACCACAAAGAUCAUCAUGAGCACAUCAAUGGCAGUGCUGGUGGC CAUGAUCUUGGGAGGAUUCUCAAUGAGUGACCUGGCUAAGCUUGUG AUCCUGAUGGGGGCCACUUUCGCAGAAAUGAACACUGGAGGAGACG UAGCUCACUUGGCAUUAGUAGCGGCAUUUAAAGUCAGACCAGCCUU GCUGGUCUCAUUUAUCUUCAGAGCCAACUGGACACCUCGUGAGAGC AUGCUGCUAGCCUUGGCUUCGUGUCUUCUGCAAACUGCGAUCUCCGC UCUUGAAGGCGACUUGAUGGUCCUCGUUAAUGGAUUUGCUUUGGCC UGGUUGGCAAUACGUGCAAUGGCCGUGCCACGCACUGACAACAUCG CUCUAGCAAUUCUGGCUGCUCUAACACCACUAGCCCGAGGCACACUG CUCGUGGCAUGGAGAGCGGGCCUCGCCACUUGUGGAGGGUUCAUGC UCCUCUCCCUGAAAGGGAAAGGUAGUGUGAAGAAGAACCUGCCAUU CGUCGCGGCCUUGGGAUUGACCGCUGUGAGAAUAGUGGACCCCAUU AAUGUGGUGGGACUACUGUUACUCACAAGGAGUGGGAAGCGGAGCU GGCCCCCUAGUGAAGUGCUCACUGCUGUCGGCCUGAUAUGUGCAUU GGCCGGAGGGUUUGCCAAGGCAGACAUAGAGAUGGCUGGGCCCAUG GCGGCAGUGGGCCUGCUAAUUGUCAGUUAUGUGGUCUCGGGAAAGA GUGUAGAUAUGUACAUUGAAAGAGCAGGUGACAUCACAUGGGAGAA AGACGCGGAAGUCACUGGAAACAGUCCUCGGCUUGACGUGGCACUA GAUGAGAGUGGUGAUUUCUCUCUGGUGGAGGAAGAUGGUCCACCCA UGAGAGAGAUCAUACUUAAGGUGGUCUUGAUGGCCAUCUGUGGCAU GAACCCAAUAGCCAUACCUUUUGCUGCAGGAGCGUGGUAUGUGUAU GUGAAGACUGGGAAAAGGAGUGGUGCCCUCUGGGACGUGCCUGCUC CGAAAGAAGUGAAAAAAGGAGAGACCACAGAUGGAGUGUACAGAGU GAUGACUCGCAGACUGCUGGGUUCAACACAAGUUGGAGUGGGAGUC AUGCAGGAGGGAGUCUUCCACACCAUGUGGCACGUCACAAAAGGGG CCGCAUUGAGGAGCGGUGAAGGGAGACUUGAUCCAUACUGGGGGGA UGUCAAGCAGGACUUGGUGUCAUAUUGUGGGCCUUGGAAGCUGGAC GCAGCUUGGGACGGAGUUAGUGAGGUGCAGCUUCUGGCCGUACCCC CUGGAGAGAGAGCCAGAAACAUUCAGACUCUGCCUGGAAUAUUUAA GACAAAGGAUGGGGACAUCGGAGCAGUUGCUUUGGACUAUCCUGCA GGAACCUCAGGAUCUCCGAUCCUAGACAAAUGCGGGAGAGUGAUAG GACUCUAUGGCAAUGGGGUUGUGAUCAAGAACGGAAGCUAUGUUAG UGCUAUAACCCAGGGAAAGAGGGAGGAGGAGACUCCGGUUGAGUGU UUUGAACCCUCGAUGCUGAAGAAGAAGCAGCUAACUGUCCUGGACC UGCAUCCAGGGGCUGGGAAAACCAGGAGAGUUCUUCCUGAAAUAGU CCGUGAAGCUAUAAAGAAGAGACUCCGCACGGUGAUCUUGGCACCA ACCAGGGUCGUCGCUGCUGAGAUGGAGGAAGCCCUGAGAGGACUUC CGGUGCGUUACAUGACAACAGCAGUCAAGGUCACCCAUUCUGGGAC AGAAAUCGUUGAUUUGAUGUGCCAUGCCACCUUCACUUCACGCCUA CUACAACCCAUUAGAGUCCCUAAUUACAACCUCUACAUCAUGGAUG AAGCCCAUUUCACAGACCCCUCAAGCAUAGCUGCAAGAGGAUAUAU AUCAACAAGGGUUGAGAUGGGCGAGGCAGCAGCCAUCUUUAUGACU GCCACACCACCAGGAACCCGCGAUGCGUUUCCAGAUUCCAACUCACC AAUCAUGGACACAGAAGUGGAAGUCCCAGAGAGAGCCUGGAGCUCA GGCUUUGAUUGGGUGACGGACCAUUCUGGGAAAACAGUUUGGUUCG UUCCAAGCGUGAGGAAUGGAAAUGAAAUCGCAGCCUGUCUGACAAA GGCUGGAAAGCGGGUUAUACAGCUUAGUAGGAAAACUUUUGAGACA GAGUUUCAGAAAACAAAAAAUCAAGAGUGGGACUUUGUCAUAACAA CUGACAUCUCAGAGAUGGGUGCCAACUUCAAGGCUGACCGGGUUAU AGAUUCCAGGAGAUGCCUAAAGCCAGUUAUACUUGAUGGUGAGAGA GUCAUCUUGGCUGGGCCCAUGCCUGUCACGCAUGCUAGCGCUGCUCA GAGGAGAGGACGUAUAGGCAGGAACCCCAACAAGCCUGGAGAUGAG UACAUGUAUGGAGGUGGGUGUGCGGAGACUGAUGAAGACCAUGCAC AUUGGCUUGAAGCAAGAAUGCUUCUUGACAACAUUUACCUCCAGGA UGGCCUCAUAGCCUCGCUCUAUCGACCUGAGGCCGACAAGGUAGCCG CCAUUGAGGGAGAGUUUAAGCUGAGGACAGAGCAAAGGAAGACCUU UGUGGAACUCAUGAAGAGAGGAGAUCUUCCCGUUUGGUUGGCCUAC CAGGUUGCAUCUGCCGGAAUAACUUAUACAGACAGAAGAUGGUGUU UUGAUGGCACAACCAACAACACCAUAAUGGAAGACAGUGUACCAGC AGAGGUGUGGACCAAGUAUGGAGAGAAGAGAGUGCUCAAACCAAGA UGGAUGGACGCCAGGGUCUGCUCAGAUCAUGCGGCCCUGAAGUCGU UCAAAGAAUUCGCCGCUGGGAAAAGAGGAGCGGCUUUGGGAGUAAU GGAGGCCCUGGGAACAUUACCAGGACACAUGACAGAGAGGUUUCAG GAAGCCAUUGAUAACCUCGCUGUGCUCAUGCGAGCAGAGACUGGAA GCAGGCCCUACAAGGCAGCGGCAGCCCAAUUGCCGGAGACCCUAGAG ACCAUCAUGCUUUUAGGCCUGCUGGGAACAGUAUCGCUGGGGAUCU UUUUUGUCUUGAUGAGGAACAAGGGCAUCGGGAAGAUGGGCUUUGA AAUGGUAACCCUUGGGGCCAGCGCAUGGCUCAUGUGGCUCUCAGAA AUCGAACCAGCCAGAAUUGCAUGUGUCCUUAUUGUUGUGUUUUUAU UACUGGUGGUGCUAAUACCAGAGCCAGAGAAGCAAAGAUCCCCCCA GGACAAUCAGAUGGCAAUCAUUAUUAUGGUGGCAGUGGGCCUUUUG GGGUUGAUAACUGCAAAUGAACUUGGAUGGCUGGAGAGAACAAAAA AUGACAUAGCUCAUCUGAUGGGAAAGAGAGAAGAGGGAACAACCGU GGGAUUCUCAAUGGACAUCGAUCUGCGACCAGCCUCCGCAUGGGCU AUUUAUGCCGCAUUGACAACCCUCAUCACCCCAGCCGUCCAGCACGC GGUAACUACCUCGUACAACAACUACUCCUUAAUGGCGAUGGCCACAC AAGCUGGAGUGCUGUUUGGCAUGGGCAAAGGGAUGCCAUUUUAUGC AUGGGACUUAGGAGUCCCGUUGCUAAUGAUGGGCUGCUACUCACAA CUAACACCCCUGACCCUGAUAGUAGCCAUCAUUUUGCUUGUGGCACA UUACAUGUACUUGAUCCCAGGCCUACAGGCAGCAGCAGCACGCGCUG CCCAGAAGAGAACAGCAGCCGGCAUCAUGAAGAAUCCCGUUGUGGA UGGAAUAGUGGUAACUGACAUUGACACAAUGACAAUUGACCCCCAA GUGGAGAAGAAGAUGGGACAAGUGCUACUUAUAGCAGUGGCUGUCU CCAGUGCUGUGUUGCUGCGGACCGCUUGGGGAUGGGGGGAGGCUGG AGCUUUGAUCACAGCAGCAACUUCCACCCUGUGGGAAGGCUCCCCAA ACAAAUACUGGAACUCCUCCACAGCCACCUCACUGUGCAACAUCUUC AGAGGAAGUUACUUGGCAGGAGCUUCCCUUAUUUACACAGUGACAA GAAAUGCCGGCCUGGUUAAGAGACGUGGAGGUGGAACGGGAGAAAC UCUGGGAGAGAAGUGGAAAGCCCGCCUGAAUCAGAUGUCGGCCUUG GAGUUCUACUCUUACAAAAAGUCAGGCAUCACUGAAGUAUGUAGAG AGGAGGCUCGCCGCGCCCUCAAGGAUGGAGUGGCCACAGGAGGACA UGCUGUAUCCCGAGGAAGCGCAAAACUCAGAUGGUUGGUGGAGAGA GGAUAUCUGCAGCCCUAUGGAAAGGUUGUUGAUCUCGGAUGCGGCA GAGGGGGCUGGAGUUAUUAUGCCGCCACCAUCCGCAAAGUGCAGGA GGUGAGAGGAUACACAAAGGGAGGUCCCGGUCAUGAAGAGCCCAUG CUGGUGCAAAGCUAUGGGUGGAACAUAAUUCGUCUCAAGAGUGGAG UGGACGUCUUCCACAUGGCGGCUGAGUCGUGUGACACUUUGCUGUG UGACAUAGGUGAGUCAUCAUCCAGUCCUGAAGUGGAGGAGACGCGA ACACUCAGAGUGCUCUCCAUGGUGGGGGACUGGCUUGAGAAGAGAC CAGGGGCCUUCUGCAUAAAGGUGUUAUGCCCAUACACCAGCACCAU GAUGGAGACCAUGGAGCGACUGCAACGUAGGUAUGGGGGAGGACUA GUCAGAGUGCCACUGUCCCGCAAUUCUACACAUGAGAUGUAUUGGG UCUCUGGAGCAAAAAGUAACAUCAUAAAAAGUGUGUCCACCACAAG UCAGCUCCUCCUGGGACGCAUGGAUGGGCCCAGGAGGCCAGUGAAG UAUGAGGAGGAUGUGAACCUCGGCUCAGGCACACGAGCUGUGGCAA GCUGUGCUGAGGCUCCCAACAUGAAGGUCAUUGGUAGGCGCAUUGA GAGAAUCCGUAGUGAACAUGCAGAAACAUGGUUCUUUGAUGAAAAC CAUCCAUACAGGACAUGGGCCUACCACGGGAGCUACGAAGCCCCCAC GCAAGGGUCAGCAUCUUCCCUCGUGAAUGGGGUUGUUAGACUCCUG UCAAAGCCCUGGGAUGUGGUGACUGGAGUUACAGGAAUAGCUAUGA CUGACACCACACCGUACGGCCAACAAAGAGUCUUCAAAGAAAAAGU GGACACCAGGGUGCCAGACCCUCAAGAAGGUACUCGCCAGGUAAUG AACAUGGUCGCUUCCUGGCUGUGGAAGGAGCUGGGAAAACGUAAGC GGCCACGUGUCUGCACCAAAGAAGAGUUCAUCAACAAGGUGCGCAG CAAUGCAGCACUGGGAGCAAUAUUUGAAGAGGAAAAAGAAUGGAAG ACGGCUGUGGAAGCUGUGAAUGAUCCAAGGUUUUGGGCCCUAGUGG AUAAGGAAAGAGAACACCACCUGAGAGGAGAGUGCCAUAGUUGUGU GUACAACAUGAUGGGAAAAAGAGAAAAGAAGCAAGGGGAAUUCGGG AAAGCAAAAGGCAGUCGCGCCAUCUGGUACAUGUGGUUGGGAGCCA GAUUCUUGGAGUUUGAAGCCCUUGGAUUCUUGAACGAGGACCAUUG GAUGGGAAGAGAAAACUCAGGAGGUGGUGUCGAAGGGUUGGGACUG CAAAGACUUGGAUACGUUCUAGAAGAAAUGAGCCGGGCACCAGGAG GAAAGAUGUAUGCAGAUGACACCGCUGGCUGGGACACCCGCAUUAG CAAGUUUGAUUUGGAGAAUGAAGCCUUGAUUACUAACCAAAUGGAU GAAGGGCACAGAACUCUGGCGUUGGCCGUGAUUAAGUACACAUACC AAAACAAAGUGGUGAAGGUCCUCAGACCAGCUGAAGGAGGAAAAAC AGUCAUGGACAUCAUUUCAAGACAAGACCAGAGGGGGAGCGGACAA GUUGUCACUUAUGCUCUCAACACAUUUACCAACUUGGUGGUGCAGC UCAUCCGGAACAUGGAGGCUGAGGAAGUGUUAGAGAUGCAAGACUU AUGGCUGUUGAGGAAGCCAGAGAAAGUAACCAGAUGGCUGCAGAGU AGCGGAUGGGACAGACUCAAACGAAUGGCAGUCAGUGGUGAUGACU GUGUUGUAAAGCCAAUUGAUGACAGGUUUGCACACGCCCUCAGGUU CUUGAAUGAUAUGGGGAAAGUUAGGAAAGACACACAGGAAUGGAAA CCCUCAACUGGAUGGAGCAACUGGGAAGAAGUCCCGUUCUGCUCCCA CCACUUUAACAAGCUGCACCUCAAAGACGGGAGAUCCAUUGUGGUC CCUUGCCGCCACCAAGAUGAACUGAUUGGCCGGGCUCGCGUUUCGCC GGGGGCAGGAUGGAGCAUCCGGGAGACUGCCUGUCUUGCAAAAUCA UAUGCACAGAUGUGGCAGCUUCUUUAUUUCCACAGAAGAGACCUCC GACUGAUGGCCAAUGCCAUUUGCUCGGCCGUGCCAGUUGACUGGGU CCCAACUGGGAGAACUACCUGGUCAAUCCAUGGAAAGGGAGAAUGG AUGACUACUGAGGACAUGCUCAUGGUGUGGAAUAGAGUGUGGAUUG AGGAGAAUGAUCACAUGGAGGACAAGACCCCUGUAACAAAAUGGAC AGACAUUCCCUAUUUGGGAAAAAGGGAGGACUUAUGGUGUGGAUCC CUUAUAGGACACAGACCUCGCACCACUUGGGCUGAGAACAUCAAAG ACACAGUCAGCAUGGUGCGCAGAAUCAUAGGUGAUGAAGAAAAGUA CAUGGACUACCUAUCCACUCAAGUUCGCUACUUGGGUGAGGAAGGG UCUACACCUGGAGUGCUGUAA IgE HC signal UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAG 138 peptide_prM-E #1 GGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACC (Brazil_isolate_ AUGGACUGGACCUGGAUCCUGUUCCUGGUGGCCGCUGCCACAAGAG ZikaSPH2015, UGCACAGCGUGGAAGUGACCAGACGGGGCAGCGCCUACUACAUGUA Sequence, NT (5′ CCUGGACAGAAGCGACGCCGGCGAGGCCAUCAGCUUUCCAACCACCC UTR, ORF, 3′ UGGGCAUGAACAAGUGCUACAUCCAGAUCAUGGACCUGGGCCACAU UTR) GUGCGACGCCACCAUGAGCUACGAGUGCCCCAUGCUGGACGAGGGCG UGGAACCCGACGAUGUGGACUGCUGGUGCAACACCACCAGCACCUGG GUGGUGUACGGCACCUGUCACCACAAGAAGGGCGAAGCCAGACGGU CCAGACGGGCCGUGACACUGCCUAGCCACAGCACCAGAAAGCUGCAG ACCCGGUCCCAGACCUGGCUGGAAAGCAGAGAGUACACCAAGCACCU GAUCCGGGUGGAAAACUGGAUCUUCCGGAACCCCGGCUUUGCCCUG GCUGCCGCUGCUAUUGCUUGGCUGCUGGGCAGCAGCACCUCCCAGAA AGUGAUCUACCUCGUGAUGAUCCUGCUGAUCGCCCCUGCCUACAGCA UCCGGUGUAUCGGCGUGUCCAACCGGGACUUCGUGGAAGGCAUGAG CGGCGGCACAUGGGUGGACGUGGUGCUGGAACAUGGCGGCUGCGUG ACAGUGAUGGCCCAGGAUAAGCCCGCCGUGGACAUCGAGCUCGUGA CCACCACCGUGUCCAAUAUGGCCGAAGUGCGGAGCUACUGCUACGAG GCCAGCAUCAGCGACAUGGCCAGCGACAGCAGAUGCCCUACACAGGG CGAGGCCUACCUGGAUAAGCAGUCCGACACCCAGUACGUGUGCAAGC GGACCCUGGUGGAUAGAGGCUGGGGCAAUGGCUGCGGCCUGUUUGG CAAGGGCAGCCUCGUGACCUGCGCCAAGUUCGCCUGCAGCAAGAAGA UGACCGGCAAGAGCAUCCAGCCCGAGAACCUGGAAUACCGGAUCAU GCUGAGCGUGCACGGCUCCCAGCACAGCGGCAUGAUCGUGAACGACA CCGGCCACGAGACAGACGAGAACCGGGCCAAGGUGGAAAUCACCCCC AACAGCCCUAGAGCCGAGGCCACACUGGGCGGCUUUGGAUCUCUGG GCCUGGACUGCGAGCCUAGAACCGGCCUGGAUUUCAGCGACCUGUAC UACCUGACCAUGAACAACAAGCACUGGCUGGUGCACAAAGAGUGGU UCCACGACAUCCCCCUGCCCUGGCAUGCUGGCGCUGAUACAGGCACC CCCCACUGGAACAACAAAGAGGCUCUGGUGGAAUUCAAGGACGCCC ACGCCAAGCGGCAGACCGUGGUGGUGCUGGGAUCUCAGGAAGGCGC CGUGCAUACAGCUCUGGCUGGCGCCCUGGAAGCCGAAAUGGAUGGC GCCAAAGGCAGACUGUCCUCCGGCCACCUGAAGUGCCGGCUGAAGAU GGACAAGCUGCGGCUGAAGGGCGUGUCCUACAGCCUGUGUACCGCC GCCUUCACCUUCACCAAGAUCCCCGCCGAGACACUGCACGGCACCGU GACUGUGGAAGUGCAGUACGCCGGCACCGACGGCCCUUGUAAAGUG CCUGCUCAGAUGGCCGUGGAUAUGCAGACCCUGACCCCCGUGGGCAG ACUGAUCACCGCCAACCCUGUGAUCACCGAGAGCACCGAGAACAGCA AGAUGAUGCUGGAACUGGACCCCCCCUUCGGCGACUCCUACAUCGUG AUCGGCGUGGGAGAGAAGAAGAUCACCCACCACUGGCACAGAUCCG GCAGCACCAUCGGCAAGGCCUUUGAGGCUACAGUGCGGGGAGCCAA GAGAAUGGCCGUGCUGGGCGAUACCGCCUGGGAUUUUGGCUCUGUG GGCGGAGCCCUGAACAGCCUGGGAAAGGGCAUCCACCAGAUCUUCG GCGCUGCCUUCAAGAGCCUGUUCGGCGGCAUGAGCUGGUUCAGCCA GAUCCUGAUCGGCACCCUGCUCGUGUGGCUGGGCCUGAACACCAAGA ACGGCAGCAUCUCCCUGACCUGCCUGGCUCUGGGCGGCGUGCUGAUC UUUCUGAGCACAGCCGUGUCCGCCUGAUAAUAGGCUGGAGCCUCGG UGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCC UUCCUGCACCCGUACCCCCGUGGUCUUUGAAUAAAGUCUGAGUGGG CGGC IgE HC signal AUGGACUGGACCUGGAUCCUGUUCCUGGUGGCCGCUGCCACAAGAG 139 peptide_prM-E #1 UGCACAGCGUGGAAGUGACCAGACGGGGCAGCGCCUACUACAUGUA (Brazil_isolate_ CCUGGACAGAAGCGACGCCGGCGAGGCCAUCAGCUUUCCAACCACCC ZikaSPH2015), ORF UGGGCAUGAACAAGUGCUACAUCCAGAUCAUGGACCUGGGCCACAU Sequence, NT GUGCGACGCCACCAUGAGCUACGAGUGCCCCAUGCUGGACGAGGGCG UGGAACCCGACGAUGUGGACUGCUGGUGCAACACCACCAGCACCUGG GUGGUGUACGGCACCUGUCACCACAAGAAGGGCGAAGCCAGACGGU CCAGACGGGCCGUGACACUGCCUAGCCACAGCACCAGAAAGCUGCAG ACCCGGUCCCAGACCUGGCUGGAAAGCAGAGAGUACACCAAGCACCU GAUCCGGGUGGAAAACUGGAUCUUCCGGAACCCCGGCUUUGCCCUG GCUGCCGCUGCUAUUGCUUGGCUGCUGGGCAGCAGCACCUCCCAGAA AGUGAUCUACCUCGUGAUGAUCCUGCUGAUCGCCCCUGCCUACAGCA UCCGGUGUAUCGGCGUGUCCAACCGGGACUUCGUGGAAGGCAUGAG CGGCGGCACAUGGGUGGACGUGGUGCUGGAACAUGGCGGCUGCGUG ACAGUGAUGGCCCAGGAUAAGCCCGCCGUGGACAUCGAGCUCGUGA CCACCACCGUGUCCAAUAUGGCCGAAGUGCGGAGCUACUGCUACGAG GCCAGCAUCAGCGACAUGGCCAGCGACAGCAGAUGCCCUACACAGGG CGAGGCCUACCUGGAUAAGCAGUCCGACACCCAGUACGUGUGCAAGC GGACCCUGGUGGAUAGAGGCUGGGGCAAUGGCUGCGGCCUGUUUGG CAAGGGCAGCCUCGUGACCUGCGCCAAGUUCGCCUGCAGCAAGAAGA UGACCGGCAAGAGCAUCCAGCCCGAGAACCUGGAAUACCGGAUCAU GCUGAGCGUGCACGGCUCCCAGCACAGCGGCAUGAUCGUGAACGACA CCGGCCACGAGACAGACGAGAACCGGGCCAAGGUGGAAAUCACCCCC AACAGCCCUAGAGCCGAGGCCACACUGGGCGGCUUUGGAUCUCUGG GCCUGGACUGCGAGCCUAGAACCGGCCUGGAUUUCAGCGACCUGUAC UACCUGACCAUGAACAACAAGCACUGGCUGGUGCACAAAGAGUGGU UCCACGACAUCCCCCUGCCCUGGCAUGCUGGCGCUGAUACAGGCACC CCCCACUGGAACAACAAAGAGGCUCUGGUGGAAUUCAAGGACGCCC ACGCCAAGCGGCAGACCGUGGUGGUGCUGGGAUCUCAGGAAGGCGC CGUGCAUACAGCUCUGGCUGGCGCCCUGGAAGCCGAAAUGGAUGGC GCCAAAGGCAGACUGUCCUCCGGCCACCUGAAGUGCCGGCUGAAGAU GGACAAGCUGCGGCUGAAGGGCGUGUCCUACAGCCUGUGUACCGCC GCCUUCACCUUCACCAAGAUCCCCGCCGAGACACUGCACGGCACCGU GACUGUGGAAGUGCAGUACGCCGGCACCGACGGCCCUUGUAAAGUG CCUGCUCAGAUGGCCGUGGAUAUGCAGACCCUGACCCCCGUGGGCAG ACUGAUCACCGCCAACCCUGUGAUCACCGAGAGCACCGAGAACAGCA AGAUGAUGCUGGAACUGGACCCCCCCUUCGGCGACUCCUACAUCGUG AUCGGCGUGGGAGAGAAGAAGAUCACCCACCACUGGCACAGAUCCG GCAGCACCAUCGGCAAGGCCUUUGAGGCUACAGUGCGGGGAGCCAA GAGAAUGGCCGUGCUGGGCGAUACCGCCUGGGAUUUUGGCUCUGUG GGCGGAGCCCUGAACAGCCUGGGAAAGGGCAUCCACCAGAUCUUCG GCGCUGCCUUCAAGAGCCUGUUCGGCGGCAUGAGCUGGUUCAGCCA GAUCCUGAUCGGCACCCUGCUCGUGUGGCUGGGCCUGAACACCAAGA ACGGCAGCAUCUCCCUGACCUGCCUGGCUCUGGGCGGCGUGCUGAUC UUUCUGAGCACAGCCGUGUCCGCC IgE HC signal G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCC 140 peptide_prM-E #1 ACCAUGGACUGGACCUGGAUCCUGUUCCUGGUGGCCGCUGCCACAAG (Brazil_isolate_ AGUGCACAGCGUGGAAGUGACCAGACGGGGCAGCGCCUACUACAUG ZikaSPH2015), mRNA UACCUGGACAGAAGCGACGCCGGCGAGGCCAUCAGCUUUCCAACCAC Sequence (T100 CCUGGGCAUGAACAAGUGCUACAUCCAGAUCAUGGACCUGGGCCAC tail) AUGUGCGACGCCACCAUGAGCUACGAGUGCCCCAUGCUGGACGAGG GCGUGGAACCCGACGAUGUGGACUGCUGGUGCAACACCACCAGCACC UGGGUGGUGUACGGCACCUGUCACCACAAGAAGGGCGAAGCCAGAC GGUCCAGACGGGCCGUGACACUGCCUAGCCACAGCACCAGAAAGCUG CAGACCCGGUCCCAGACCUGGCUGGAAAGCAGAGAGUACACCAAGCA CCUGAUCCGGGUGGAAAACUGGAUCUUCCGGAACCCCGGCUUUGCCC UGGCUGCCGCUGCUAUUGCUUGGCUGCUGGGCAGCAGCACCUCCCAG AAAGUGAUCUACCUCGUGAUGAUCCUGCUGAUCGCCCCUGCCUACAG CAUCCGGUGUAUCGGCGUGUCCAACCGGGACUUCGUGGAAGGCAUG AGCGGCGGCACAUGGGUGGACGUGGUGCUGGAACAUGGCGGCUGCG UGACAGUGAUGGCCCAGGAUAAGCCCGCCGUGGACAUCGAGCUCGU GACCACCACCGUGUCCAAUAUGGCCGAAGUGCGGAGCUACUGCUACG AGGCCAGCAUCAGCGACAUGGCCAGCGACAGCAGAUGCCCUACACAG GGCGAGGCCUACCUGGAUAAGCAGUCCGACACCCAGUACGUGUGCA AGCGGACCCUGGUGGAUAGAGGCUGGGGCAAUGGCUGCGGCCUGUU UGGCAAGGGCAGCCUCGUGACCUGCGCCAAGUUCGCCUGCAGCAAGA AGAUGACCGGCAAGAGCAUCCAGCCCGAGAACCUGGAAUACCGGAU CAUGCUGAGCGUGCACGGCUCCCAGCACAGCGGCAUGAUCGUGAACG ACACCGGCCACGAGACAGACGAGAACCGGGCCAAGGUGGAAAUCACC CCCAACAGCCCUAGAGCCGAGGCCACACUGGGCGGCUUUGGAUCUCU GGGCCUGGACUGCGAGCCUAGAACCGGCCUGGAUUUCAGCGACCUG UACUACCUGACCAUGAACAACAAGCACUGGCUGGUGCACAAAGAGU GGUUCCACGACAUCCCCCUGCCCUGGCAUGCUGGCGCUGAUACAGGC ACCCCCCACUGGAACAACAAAGAGGCUCUGGUGGAAUUCAAGGACG CCCACGCCAAGCGGCAGACCGUGGUGGUGCUGGGAUCUCAGGAAGG CGCCGUGCAUACAGCUCUGGCUGGCGCCCUGGAAGCCGAAAUGGAU GGCGCCAAAGGCAGACUGUCCUCCGGCCACCUGAAGUGCCGGCUGAA GAUGGACAAGCUGCGGCUGAAGGGCGUGUCCUACAGCCUGUGUACC GCCGCCUUCACCUUCACCAAGAUCCCCGCCGAGACACUGCACGGCAC CGUGACUGUGGAAGUGCAGUACGCCGGCACCGACGGCCCUUGUAAA GUGCCUGCUCAGAUGGCCGUGGAUAUGCAGACCCUGACCCCCGUGGG CAGACUGAUCACCGCCAACCCUGUGAUCACCGAGAGCACCGAGAACA GCAAGAUGAUGCUGGAACUGGACCCCCCCUUCGGCGACUCCUACAUC GUGAUCGGCGUGGGAGAGAAGAAGAUCACCCACCACUGGCACAGAU CCGGCAGCACCAUCGGCAAGGCCUUUGAGGCUACAGUGCGGGGAGCC AAGAGAAUGGCCGUGCUGGGCGAUACCGCCUGGGAUUUUGGCUCUG UGGGCGGAGCCCUGAACAGCCUGGGAAAGGGCAUCCACCAGAUCUU CGGCGCUGCCUUCAAGAGCCUGUUCGGCGGCAUGAGCUGGUUCAGCC AGAUCCUGAUCGGCACCCUGCUCGUGUGGCUGGGCCUGAACACCAAG AACGGCAGCAUCUCCCUGACCUGCCUGGCUCUGGGCGGCGUGCUGAU CUUUCUGAGCACAGCCGUGUCCGCCUGAUAAUAGGCUGGAGCCUCG GUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCC CUUCCUGCACCCGUACCCCCGUGGUCUUUGAAUAAAGUCUGAGUGG GCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAUCUAG IgE HC signal UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAG 141 peptide_prM-E #2 GGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACC (Brazil_isolate_ AUGGACUGGACCUGGAUCCUGUUCCUGGUGGCCGCUGCCACAAGAG ZikaSPH2015), UGCACAGCACCAGAAGAGGCAGCGCCUACUACAUGUACCUGGACAG Sequence, NT (5′ AAGCGACGCCGGCGAGGCCAUCAGCUUUCCAACCACCCUGGGCAUGA UTR, ORF, 3′ ACAAGUGCUACAUCCAGAUCAUGGACCUGGGCCACAUGUGCGACGCC UTR) ACCAUGAGCUACGAGUGCCCCAUGCUGGACGAGGGCGUGGAACCCG ACGAUGUGGACUGCUGGUGCAACACCACCAGCACCUGGGUGGUGUA CGGCACCUGUCACCACAAGAAGGGCGAAGCCAGACGGUCCAGACGGG CCGUGACACUGCCUAGCCACUCCACCAGAAAGCUGCAGACCCGGUCC CAGACCUGGCUGGAAAGCAGAGAGUACACCAAGCACCUGAUCCGGG UGGAAAACUGGAUCUUCCGGAACCCCGGCUUUGCCCUGGCUGCCGCU GCUAUUGCUUGGCUGCUGGGCAGCAGCACCUCCCAGAAAGUGAUCU ACCUCGUGAUGAUCCUGCUGAUCGCCCCUGCCUACAGCAUCCGGUGU AUCGGCGUGUCCAACCGGGACUUCGUGGAAGGCAUGAGCGGCGGCA CAUGGGUGGACGUGGUGCUGGAACAUGGCGGCUGCGUGACAGUGAU GGCCCAGGAUAAGCCCGCCGUGGACAUCGAGCUCGUGACCACCACCG UGUCCAAUAUGGCCGAAGUGCGGAGCUACUGCUACGAGGCCAGCAU CAGCGACAUGGCCAGCGACAGCAGAUGCCCUACACAGGGCGAGGCCU ACCUGGAUAAGCAGUCCGACACCCAGUACGUGUGCAAGCGGACCCUG GUGGAUAGAGGCUGGGGCAAUGGCUGCGGCCUGUUUGGCAAGGGCA GCCUCGUGACCUGCGCCAAGUUCGCCUGCAGCAAGAAGAUGACCGGC AAGAGCAUCCAGCCCGAGAACCUGGAAUACCGGAUCAUGCUGAGCG UGCACGGCUCCCAGCACAGCGGCAUGAUCGUGAACGACACCGGCCAC GAGACAGACGAGAACCGGGCCAAGGUGGAAAUCACCCCCAACAGCCC UAGAGCCGAGGCCACACUGGGCGGCUUUGGAUCUCUGGGCCUGGAC UGCGAGCCUAGAACCGGCCUGGAUUUCAGCGACCUGUACUACCUGAC CAUGAACAACAAGCACUGGCUGGUGCACAAAGAGUGGUUCCACGAC AUCCCCCUGCCCUGGCAUGCUGGCGCUGAUACAGGCACCCCCCACUG GAACAACAAAGAGGCUCUGGUGGAAUUCAAGGACGCCCACGCCAAG CGGCAGACCGUGGUGGUGCUGGGAUCUCAGGAAGGCGCCGUGCAUA CAGCUCUGGCUGGCGCCCUGGAAGCCGAAAUGGAUGGCGCCAAAGG CAGACUGUCCUCCGGCCACCUGAAGUGCCGGCUGAAGAUGGACAAGC UGCGGCUGAAGGGCGUGUCCUACAGCCUGUGUACCGCCGCCUUCACC UUCACCAAGAUCCCCGCCGAGACACUGCACGGCACCGUGACUGUGGA AGUGCAGUACGCCGGCACCGACGGCCCUUGUAAAGUGCCUGCUCAGA UGGCCGUGGAUAUGCAGACCCUGACCCCCGUGGGCAGACUGAUCACC GCCAACCCUGUGAUCACCGAGAGCACCGAGAACAGCAAGAUGAUGC UGGAACUGGACCCCCCCUUCGGCGACUCCUACAUCGUGAUCGGCGUG GGAGAGAAGAAGAUCACCCACCACUGGCACAGAUCCGGCAGCACCAU CGGCAAGGCCUUUGAGGCUACAGUGCGGGGAGCCAAGAGAAUGGCC GUGCUGGGCGAUACCGCCUGGGAUUUUGGCUCUGUGGGCGGAGCCC UGAACAGCCUGGGAAAGGGCAUCCACCAGAUCUUCGGAGCCGCCUU UAAGAGCCUGUUCGGCGGCAUGAGCUGGUUCAGCCAGAUCCUGAUC GGCACCCUGCUCGUGUGGCUGGGCCUGAACACCAAGAACGGCAGCAU CUCCCUGACCUGCCUGGCUCUGGGCGGCGUGCUGAUCUUUCUGAGCA CAGCCGUGUCCGCCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCU UCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCC GUACCCCCGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC IgE HC signal AUGGACUGGACCUGGAUCCUGUUCCUGGUGGCCGCUGCCACAAGAG 142 peptide_prM-E #2 UGCACAGCACCAGAAGAGGCAGCGCCUACUACAUGUACCUGGACAG (Brazil_isolate_ AAGCGACGCCGGCGAGGCCAUCAGCUUUCCAACCACCCUGGGCAUGA ZikaSPH2015), ORF ACAAGUGCUACAUCCAGAUCAUGGACCUGGGCCACAUGUGCGACGCC Sequence, NT ACCAUGAGCUACGAGUGCCCCAUGCUGGACGAGGGCGUGGAACCCG ACGAUGUGGACUGCUGGUGCAACACCACCAGCACCUGGGUGGUGUA CGGCACCUGUCACCACAAGAAGGGCGAAGCCAGACGGUCCAGACGGG CCGUGACACUGCCUAGCCACUCCACCAGAAAGCUGCAGACCCGGUCC CAGACCUGGCUGGAAAGCAGAGAGUACACCAAGCACCUGAUCCGGG UGGAAAACUGGAUCUUCCGGAACCCCGGCUUUGCCCUGGCUGCCGCU GCUAUUGCUUGGCUGCUGGGCAGCAGCACCUCCCAGAAAGUGAUCU ACCUCGUGAUGAUCCUGCUGAUCGCCCCUGCCUACAGCAUCCGGUGU AUCGGCGUGUCCAACCGGGACUUCGUGGAAGGCAUGAGCGGCGGCA CAUGGGUGGACGUGGUGCUGGAACAUGGCGGCUGCGUGACAGUGAU GGCCCAGGAUAAGCCCGCCGUGGACAUCGAGCUCGUGACCACCACCG UGUCCAAUAUGGCCGAAGUGCGGAGCUACUGCUACGAGGCCAGCAU CAGCGACAUGGCCAGCGACAGCAGAUGCCCUACACAGGGCGAGGCCU ACCUGGAUAAGCAGUCCGACACCCAGUACGUGUGCAAGCGGACCCUG GUGGAUAGAGGCUGGGGCAAUGGCUGCGGCCUGUUUGGCAAGGGCA GCCUCGUGACCUGCGCCAAGUUCGCCUGCAGCAAGAAGAUGACCGGC AAGAGCAUCCAGCCCGAGAACCUGGAAUACCGGAUCAUGCUGAGCG UGCACGGCUCCCAGCACAGCGGCAUGAUCGUGAACGACACCGGCCAC GAGACAGACGAGAACCGGGCCAAGGUGGAAAUCACCCCCAACAGCCC UAGAGCCGAGGCCACACUGGGCGGCUUUGGAUCUCUGGGCCUGGAC UGCGAGCCUAGAACCGGCCUGGAUUUCAGCGACCUGUACUACCUGAC CAUGAACAACAAGCACUGGCUGGUGCACAAAGAGUGGUUCCACGAC AUCCCCCUGCCCUGGCAUGCUGGCGCUGAUACAGGCACCCCCCACUG GAACAACAAAGAGGCUCUGGUGGAAUUCAAGGACGCCCACGCCAAG CGGCAGACCGUGGUGGUGCUGGGAUCUCAGGAAGGCGCCGUGCAUA CAGCUCUGGCUGGCGCCCUGGAAGCCGAAAUGGAUGGCGCCAAAGG CAGACUGUCCUCCGGCCACCUGAAGUGCCGGCUGAAGAUGGACAAGC UGCGGCUGAAGGGCGUGUCCUACAGCCUGUGUACCGCCGCCUUCACC UUCACCAAGAUCCCCGCCGAGACACUGCACGGCACCGUGACUGUGGA AGUGCAGUACGCCGGCACCGACGGCCCUUGUAAAGUGCCUGCUCAGA UGGCCGUGGAUAUGCAGACCCUGACCCCCGUGGGCAGACUGAUCACC GCCAACCCUGUGAUCACCGAGAGCACCGAGAACAGCAAGAUGAUGC UGGAACUGGACCCCCCCUUCGGCGACUCCUACAUCGUGAUCGGCGUG GGAGAGAAGAAGAUCACCCACCACUGGCACAGAUCCGGCAGCACCAU CGGCAAGGCCUUUGAGGCUACAGUGCGGGGAGCCAAGAGAAUGGCC GUGCUGGGCGAUACCGCCUGGGAUUUUGGCUCUGUGGGCGGAGCCC UGAACAGCCUGGGAAAGGGCAUCCACCAGAUCUUCGGAGCCGCCUU UAAGAGCCUGUUCGGCGGCAUGAGCUGGUUCAGCCAGAUCCUGAUC GGCACCCUGCUCGUGUGGCUGGGCCUGAACACCAAGAACGGCAGCAU CUCCCUGACCUGCCUGGCUCUGGGCGGCGUGCUGAUCUUUCUGAGCA CAGCCGUGUCCGCC IgE HC signal G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCC 143 peptide_prM-E #2 ACCAUGGACUGGACCUGGAUCCUGUUCCUGGUGGCCGCUGCCACAAG (Brazil_isolate_ AGUGCACAGCACCAGAAGAGGCAGCGCCUACUACAUGUACCUGGAC ZikaSPH2015), mRNA AGAAGCGACGCCGGCGAGGCCAUCAGCUUUCCAACCACCCUGGGCAU Sequence (T100 GAACAAGUGCUACAUCCAGAUCAUGGACCUGGGCCACAUGUGCGAC tail) GCCACCAUGAGCUACGAGUGCCCCAUGCUGGACGAGGGCGUGGAACC CGACGAUGUGGACUGCUGGUGCAACACCACCAGCACCUGGGUGGUG UACGGCACCUGUCACCACAAGAAGGGCGAAGCCAGACGGUCCAGACG GGCCGUGACACUGCCUAGCCACUCCACCAGAAAGCUGCAGACCCGGU CCCAGACCUGGCUGGAAAGCAGAGAGUACACCAAGCACCUGAUCCGG GUGGAAAACUGGAUCUUCCGGAACCCCGGCUUUGCCCUGGCUGCCGC UGCUAUUGCUUGGCUGCUGGGCAGCAGCACCUCCCAGAAAGUGAUC UACCUCGUGAUGAUCCUGCUGAUCGCCCCUGCCUACAGCAUCCGGUG UAUCGGCGUGUCCAACCGGGACUUCGUGGAAGGCAUGAGCGGCGGC ACAUGGGUGGACGUGGUGCUGGAACAUGGCGGCUGCGUGACAGUGA UGGCCCAGGAUAAGCCCGCCGUGGACAUCGAGCUCGUGACCACCACC GUGUCCAAUAUGGCCGAAGUGCGGAGCUACUGCUACGAGGCCAGCA UCAGCGACAUGGCCAGCGACAGCAGAUGCCCUACACAGGGCGAGGCC UACCUGGAUAAGCAGUCCGACACCCAGUACGUGUGCAAGCGGACCCU GGUGGAUAGAGGCUGGGGCAAUGGCUGCGGCCUGUUUGGCAAGGGC AGCCUCGUGACCUGCGCCAAGUUCGCCUGCAGCAAGAAGAUGACCGG CAAGAGCAUCCAGCCCGAGAACCUGGAAUACCGGAUCAUGCUGAGC GUGCACGGCUCCCAGCACAGCGGCAUGAUCGUGAACGACACCGGCCA CGAGACAGACGAGAACCGGGCCAAGGUGGAAAUCACCCCCAACAGCC CUAGAGCCGAGGCCACACUGGGCGGCUUUGGAUCUCUGGGCCUGGA CUGCGAGCCUAGAACCGGCCUGGAUUUCAGCGACCUGUACUACCUGA CCAUGAACAACAAGCACUGGCUGGUGCACAAAGAGUGGUUCCACGA CAUCCCCCUGCCCUGGCAUGCUGGCGCUGAUACAGGCACCCCCCACU GGAACAACAAAGAGGCUCUGGUGGAAUUCAAGGACGCCCACGCCAA GCGGCAGACCGUGGUGGUGCUGGGAUCUCAGGAAGGCGCCGUGCAU ACAGCUCUGGCUGGCGCCCUGGAAGCCGAAAUGGAUGGCGCCAAAG GCAGACUGUCCUCCGGCCACCUGAAGUGCCGGCUGAAGAUGGACAA GCUGCGGCUGAAGGGCGUGUCCUACAGCCUGUGUACCGCCGCCUUCA CCUUCACCAAGAUCCCCGCCGAGACACUGCACGGCACCGUGACUGUG GAAGUGCAGUACGCCGGCACCGACGGCCCUUGUAAAGUGCCUGCUCA GAUGGCCGUGGAUAUGCAGACCCUGACCCCCGUGGGCAGACUGAUC ACCGCCAACCCUGUGAUCACCGAGAGCACCGAGAACAGCAAGAUGAU GCUGGAACUGGACCCCCCCUUCGGCGACUCCUACAUCGUGAUCGGCG UGGGAGAGAAGAAGAUCACCCACCACUGGCACAGAUCCGGCAGCACC AUCGGCAAGGCCUUUGAGGCUACAGUGCGGGGAGCCAAGAGAAUGG CCGUGCUGGGCGAUACCGCCUGGGAUUUUGGCUCUGUGGGCGGAGC CCUGAACAGCCUGGGAAAGGGCAUCCACCAGAUCUUCGGAGCCGCCU UUAAGAGCCUGUUCGGCGGCAUGAGCUGGUUCAGCCAGAUCCUGAU CGGCACCCUGCUCGUGUGGCUGGGCCUGAACACCAAGAACGGCAGCA UCUCCCUGACCUGCCUGGCUCUGGGCGGCGUGCUGAUCUUUCUGAGC ACAGCCGUGUCCGCCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGC UUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACC CGUACCCCCGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGCAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAUCUAG HuIgG_(k) signal UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAG 144 peptide_prME #1 GGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACC (Brazil_isolate_ AUGGAAACCCCUGCCCAGCUGCUGUUCCUGCUGCUGCUGUGGCUGCC ZikaSPH2015), UGAUACCACCGGCGUGGAAGUGACCAGAAGAGGCAGCGCCUACUAC Sequence, NT (5′ AUGUACCUGGACAGAAGCGACGCCGGCGAGGCCAUCAGCUUUCCAAC UTR, ORF, 3′ CACCCUGGGCAUGAACAAGUGCUACAUCCAGAUCAUGGACCUGGGCC UTR) ACAUGUGCGACGCCACCAUGAGCUACGAGUGCCCCAUGCUGGACGAG GGCGUGGAACCCGACGAUGUGGACUGCUGGUGCAACACCACCAGCAC CUGGGUGGUGUACGGCACCUGUCACCACAAGAAGGGCGAAGCCAGA CGGUCCAGACGGGCCGUGACACUGCCUAGCCACUCCACCAGAAAGCU GCAGACCCGGUCCCAGACCUGGCUGGAAAGCAGAGAGUACACCAAGC ACCUGAUCCGGGUGGAAAACUGGAUCUUCCGGAACCCCGGCUUUGCC CUGGCCGCUGCUGCUAUUGCUUGGCUGCUGGGCAGCAGCACCUCCCA GAAAGUGAUCUACCUCGUGAUGAUCCUGCUGAUCGCCCCUGCCUACA GCAUCCGGUGUAUCGGCGUGUCCAACCGGGACUUCGUGGAAGGCAU GAGCGGCGGCACAUGGGUGGACGUGGUGCUGGAACAUGGCGGCUGC GUGACAGUGAUGGCCCAGGAUAAGCCCGCCGUGGACAUCGAGCUCG UGACCACCACCGUGUCCAAUAUGGCCGAAGUGCGGAGCUACUGCUAC GAGGCCAGCAUCAGCGACAUGGCCAGCGACAGCAGAUGCCCUACACA GGGCGAGGCCUACCUGGAUAAGCAGUCCGACACCCAGUACGUGUGC AAGCGGACCCUGGUGGAUAGAGGCUGGGGCAAUGGCUGCGGCCUGU UUGGCAAGGGCAGCCUCGUGACCUGCGCCAAGUUCGCCUGCAGCAAG AAGAUGACCGGCAAGAGCAUCCAGCCCGAGAACCUGGAAUACCGGA UCAUGCUGAGCGUGCACGGCAGCCAGCACUCCGGCAUGAUCGUGAAC GACACCGGCCACGAGACAGACGAGAACCGGGCCAAGGUGGAAAUCA CCCCCAACAGCCCUAGAGCCGAGGCCACACUGGGCGGCUUUGGAUCU CUGGGCCUGGACUGCGAGCCUAGAACCGGCCUGGAUUUCAGCGACCU GUACUACCUGACCAUGAACAACAAGCACUGGCUGGUGCACAAAGAG UGGUUCCACGACAUCCCCCUGCCCUGGCAUGCCGGCGCUGAUACAGG CACACCCCACUGGAACAACAAAGAGGCUCUGGUGGAAUUCAAGGAC GCCCACGCCAAGCGGCAGACCGUGGUGGUGCUGGGAUCUCAGGAAG GCGCCGUGCAUACAGCUCUGGCUGGCGCCCUGGAAGCCGAAAUGGA UGGCGCCAAAGGCAGACUGUCCAGCGGCCACCUGAAGUGCCGGCUGA AGAUGGACAAGCUGCGGCUGAAGGGCGUGUCCUACAGCCUGUGUAC CGCCGCCUUCACCUUCACCAAGAUCCCCGCCGAGACACUGCACGGCA CCGUGACUGUGGAAGUGCAGUACGCCGGCACCGACGGCCCUUGUAA AGUGCCUGCUCAGAUGGCCGUGGAUAUGCAGACCCUGACCCCCGUGG GCAGACUGAUCACCGCCAACCCUGUGAUCACCGAGAGCACCGAGAAC AGCAAGAUGAUGCUGGAACUGGACCCCCCCUUCGGCGACUCCUACAU CGUGAUCGGCGUGGGAGAGAAGAAGAUCACCCACCACUGGCACCGC AGCGGCAGCACAAUCGGCAAGGCCUUUGAAGCCACAGUGCGGGGAG CCAAGAGAAUGGCCGUGCUGGGAGAUACCGCCUGGGACUUUGGCUC UGUGGGCGGAGCCCUGAACUCUCUGGGCAAGGGAAUCCACCAGAUC UUCGGAGCCGCCUUUAAGAGCCUGUUCGGCGGCAUGAGCUGGUUCA GCCAGAUCCUGAUCGGCACCCUGCUCGUGUGGCUGGGCCUGAACACC AAGAACGGCAGCAUCUCCCUGACCUGCCUGGCUCUGGGAGGCGUGCU GAUCUUUCUGAGCACCGCCGUGUCUGCCUGAUAAUAGGCUGGAGCC UCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCU CCCCUUCCUGCACCCGUACCCCCGUGGUCUUUGAAUAAAGUCUGAGU GGGCGGC HuIgG_(k) signal AUGGAAACCCCUGCCCAGCUGCUGUUCCUGCUGCUGCUGUGGCUGCC 145 peptide_prME #1 UGAUACCACCGGCGUGGAAGUGACCAGAAGAGGCAGCGCCUACUAC (Brazil_isolate_ AUGUACCUGGACAGAAGCGACGCCGGCGAGGCCAUCAGCUUUCCAAC ZikaSPH2015), ORF CACCCUGGGCAUGAACAAGUGCUACAUCCAGAUCAUGGACCUGGGCC Sequence, NT ACAUGUGCGACGCCACCAUGAGCUACGAGUGCCCCAUGCUGGACGAG GGCGUGGAACCCGACGAUGUGGACUGCUGGUGCAACACCACCAGCAC CUGGGUGGUGUACGGCACCUGUCACCACAAGAAGGGCGAAGCCAGA CGGUCCAGACGGGCCGUGACACUGCCUAGCCACUCCACCAGAAAGCU GCAGACCCGGUCCCAGACCUGGCUGGAAAGCAGAGAGUACACCAAGC ACCUGAUCCGGGUGGAAAACUGGAUCUUCCGGAACCCCGGCUUUGCC CUGGCCGCUGCUGCUAUUGCUUGGCUGCUGGGCAGCAGCACCUCCCA GAAAGUGAUCUACCUCGUGAUGAUCCUGCUGAUCGCCCCUGCCUACA GCAUCCGGUGUAUCGGCGUGUCCAACCGGGACUUCGUGGAAGGCAU GAGCGGCGGCACAUGGGUGGACGUGGUGCUGGAACAUGGCGGCUGC GUGACAGUGAUGGCCCAGGAUAAGCCCGCCGUGGACAUCGAGCUCG UGACCACCACCGUGUCCAAUAUGGCCGAAGUGCGGAGCUACUGCUAC GAGGCCAGCAUCAGCGACAUGGCCAGCGACAGCAGAUGCCCUACACA GGGCGAGGCCUACCUGGAUAAGCAGUCCGACACCCAGUACGUGUGC AAGCGGACCCUGGUGGAUAGAGGCUGGGGCAAUGGCUGCGGCCUGU UUGGCAAGGGCAGCCUCGUGACCUGCGCCAAGUUCGCCUGCAGCAAG AAGAUGACCGGCAAGAGCAUCCAGCCCGAGAACCUGGAAUACCGGA UCAUGCUGAGCGUGCACGGCAGCCAGCACUCCGGCAUGAUCGUGAAC GACACCGGCCACGAGACAGACGAGAACCGGGCCAAGGUGGAAAUCA CCCCCAACAGCCCUAGAGCCGAGGCCACACUGGGCGGCUUUGGAUCU CUGGGCCUGGACUGCGAGCCUAGAACCGGCCUGGAUUUCAGCGACCU GUACUACCUGACCAUGAACAACAAGCACUGGCUGGUGCACAAAGAG UGGUUCCACGACAUCCCCCUGCCCUGGCAUGCCGGCGCUGAUACAGG CACACCCCACUGGAACAACAAAGAGGCUCUGGUGGAAUUCAAGGAC GCCCACGCCAAGCGGCAGACCGUGGUGGUGCUGGGAUCUCAGGAAG GCGCCGUGCAUACAGCUCUGGCUGGCGCCCUGGAAGCCGAAAUGGA UGGCGCCAAAGGCAGACUGUCCAGCGGCCACCUGAAGUGCCGGCUGA AGAUGGACAAGCUGCGGCUGAAGGGCGUGUCCUACAGCCUGUGUAC CGCCGCCUUCACCUUCACCAAGAUCCCCGCCGAGACACUGCACGGCA CCGUGACUGUGGAAGUGCAGUACGCCGGCACCGACGGCCCUUGUAA AGUGCCUGCUCAGAUGGCCGUGGAUAUGCAGACCCUGACCCCCGUGG GCAGACUGAUCACCGCCAACCCUGUGAUCACCGAGAGCACCGAGAAC AGCAAGAUGAUGCUGGAACUGGACCCCCCCUUCGGCGACUCCUACAU CGUGAUCGGCGUGGGAGAGAAGAAGAUCACCCACCACUGGCACCGC AGCGGCAGCACAAUCGGCAAGGCCUUUGAAGCCACAGUGCGGGGAG CCAAGAGAAUGGCCGUGCUGGGAGAUACCGCCUGGGACUUUGGCUC UGUGGGCGGAGCCCUGAACUCUCUGGGCAAGGGAAUCCACCAGAUC UUCGGAGCCGCCUUUAAGAGCCUGUUCGGCGGCAUGAGCUGGUUCA GCCAGAUCCUGAUCGGCACCCUGCUCGUGUGGCUGGGCCUGAACACC AAGAACGGCAGCAUCUCCCUGACCUGCCUGGCUCUGGGAGGCGUGCU GAUCUUUCUGAGCACCGCCGUGUCUGCC HuIgG_(k) signal G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCC 146 peptide_prME #1 ACCAUGGAAACCCCUGCCCAGCUGCUGUUCCUGCUGCUGCUGUGGCU (Brazil_isolate_ GCCUGAUACCACCGGCGUGGAAGUGACCAGAAGAGGCAGCGCCUAC ZikaSPH2015), mRNA UACAUGUACCUGGACAGAAGCGACGCCGGCGAGGCCAUCAGCUUUCC Sequence (T100 AACCACCCUGGGCAUGAACAAGUGCUACAUCCAGAUCAUGGACCUG tail) GGCCACAUGUGCGACGCCACCAUGAGCUACGAGUGCCCCAUGCUGGA CGAGGGCGUGGAACCCGACGAUGUGGACUGCUGGUGCAACACCACC AGCACCUGGGUGGUGUACGGCACCUGUCACCACAAGAAGGGCGAAG CCAGACGGUCCAGACGGGCCGUGACACUGCCUAGCCACUCCACCAGA AAGCUGCAGACCCGGUCCCAGACCUGGCUGGAAAGCAGAGAGUACA CCAAGCACCUGAUCCGGGUGGAAAACUGGAUCUUCCGGAACCCCGGC UUUGCCCUGGCCGCUGCUGCUAUUGCUUGGCUGCUGGGCAGCAGCAC CUCCCAGAAAGUGAUCUACCUCGUGAUGAUCCUGCUGAUCGCCCCUG CCUACAGCAUCCGGUGUAUCGGCGUGUCCAACCGGGACUUCGUGGA AGGCAUGAGCGGCGGCACAUGGGUGGACGUGGUGCUGGAACAUGGC GGCUGCGUGACAGUGAUGGCCCAGGAUAAGCCCGCCGUGGACAUCG AGCUCGUGACCACCACCGUGUCCAAUAUGGCCGAAGUGCGGAGCUAC UGCUACGAGGCCAGCAUCAGCGACAUGGCCAGCGACAGCAGAUGCCC UACACAGGGCGAGGCCUACCUGGAUAAGCAGUCCGACACCCAGUACG UGUGCAAGCGGACCCUGGUGGAUAGAGGCUGGGGCAAUGGCUGCGG CCUGUUUGGCAAGGGCAGCCUCGUGACCUGCGCCAAGUUCGCCUGCA GCAAGAAGAUGACCGGCAAGAGCAUCCAGCCCGAGAACCUGGAAUA CCGGAUCAUGCUGAGCGUGCACGGCAGCCAGCACUCCGGCAUGAUCG UGAACGACACCGGCCACGAGACAGACGAGAACCGGGCCAAGGUGGA AAUCACCCCCAACAGCCCUAGAGCCGAGGCCACACUGGGCGGCUUUG GAUCUCUGGGCCUGGACUGCGAGCCUAGAACCGGCCUGGAUUUCAG CGACCUGUACUACCUGACCAUGAACAACAAGCACUGGCUGGUGCACA AAGAGUGGUUCCACGACAUCCCCCUGCCCUGGCAUGCCGGCGCUGAU ACAGGCACACCCCACUGGAACAACAAAGAGGCUCUGGUGGAAUUCA AGGACGCCCACGCCAAGCGGCAGACCGUGGUGGUGCUGGGAUCUCA GGAAGGCGCCGUGCAUACAGCUCUGGCUGGCGCCCUGGAAGCCGAA AUGGAUGGCGCCAAAGGCAGACUGUCCAGCGGCCACCUGAAGUGCC GGCUGAAGAUGGACAAGCUGCGGCUGAAGGGCGUGUCCUACAGCCU GUGUACCGCCGCCUUCACCUUCACCAAGAUCCCCGCCGAGACACUGC ACGGCACCGUGACUGUGGAAGUGCAGUACGCCGGCACCGACGGCCCU UGUAAAGUGCCUGCUCAGAUGGCCGUGGAUAUGCAGACCCUGACCC CCGUGGGCAGACUGAUCACCGCCAACCCUGUGAUCACCGAGAGCACC GAGAACAGCAAGAUGAUGCUGGAACUGGACCCCCCCUUCGGCGACUC CUACAUCGUGAUCGGCGUGGGAGAGAAGAAGAUCACCCACCACUGG CACCGCAGCGGCAGCACAAUCGGCAAGGCCUUUGAAGCCACAGUGCG GGGAGCCAAGAGAAUGGCCGUGCUGGGAGAUACCGCCUGGGACUUU GGCUCUGUGGGCGGAGCCCUGAACUCUCUGGGCAAGGGAAUCCACC AGAUCUUCGGAGCCGCCUUUAAGAGCCUGUUCGGCGGCAUGAGCUG GUUCAGCCAGAUCCUGAUCGGCACCCUGCUCGUGUGGCUGGGCCUGA ACACCAAGAACGGCAGCAUCUCCCUGACCUGCCUGGCUCUGGGAGGC GUGCUGAUCUUUCUGAGCACCGCCGUGUCUGCCUGAUAAUAGGCUG GAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCC CUCCUCCCCUUCCUGCACCCGUACCCCCGUGGUCUUUGAAUAAAGUC UGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAUCUAG HuIgG_(k) signal UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAG 147 peptide_prME #2 GGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACC (Brazil_isolate_ AUGGAAACCCCUGCCCAGCUGCUGUUCCUGCUGCUGCUGUGGCUGCC ZikaSPH2015), UGAUACCACCGGCACCAGAAGAGGCAGCGCCUACUACAUGUACCUGG Sequence, NT (5′ ACAGAAGCGACGCCGGCGAGGCCAUCAGCUUUCCAACCACCCUGGGC UTR, ORF, 3′ AUGAACAAGUGCUACAUCCAGAUCAUGGACCUGGGCCACAUGUGCG UTR) ACGCCACCAUGAGCUACGAGUGCCCCAUGCUGGACGAGGGCGUGGA ACCCGACGAUGUGGACUGCUGGUGCAACACCACCAGCACCUGGGUGG UGUACGGCACCUGUCACCACAAGAAGGGCGAAGCCAGACGGUCCAG ACGGGCCGUGACACUGCCUAGCCACUCCACCAGAAAGCUGCAGACCC GGUCCCAGACCUGGCUGGAAAGCAGAGAGUACACCAAGCACCUGAU CCGGGUGGAAAACUGGAUCUUCCGGAACCCCGGCUUUGCCCUGGCCG CUGCUGCUAUUGCUUGGCUGCUGGGCAGCAGCACCUCCCAGAAAGU GAUCUACCUCGUGAUGAUCCUGCUGAUCGCCCCUGCCUACAGCAUCC GGUGUAUCGGCGUGUCCAACCGGGACUUCGUGGAAGGCAUGAGCGG CGGCACAUGGGUGGACGUGGUGCUGGAACAUGGCGGCUGCGUGACA GUGAUGGCCCAGGAUAAGCCCGCCGUGGACAUCGAGCUCGUGACCAC CACCGUGUCCAAUAUGGCCGAAGUGCGGAGCUACUGCUACGAGGCC AGCAUCAGCGACAUGGCCAGCGACAGCAGAUGCCCUACACAGGGCGA GGCCUACCUGGAUAAGCAGUCCGACACCCAGUACGUGUGCAAGCGG ACCCUGGUGGAUAGAGGCUGGGGCAAUGGCUGCGGCCUGUUUGGCA AGGGCAGCCUCGUGACCUGCGCCAAGUUCGCCUGCAGCAAGAAGAU GACCGGCAAGAGCAUCCAGCCCGAGAACCUGGAAUACCGGAUCAUGC UGAGCGUGCACGGCAGCCAGCACUCCGGCAUGAUCGUGAACGACACC GGCCACGAGACAGACGAGAACCGGGCCAAGGUGGAAAUCACCCCCAA CAGCCCUAGAGCCGAGGCCACACUGGGCGGCUUUGGAUCUCUGGGCC UGGACUGCGAGCCUAGAACCGGCCUGGAUUUCAGCGACCUGUACUA CCUGACCAUGAACAACAAGCACUGGCUGGUGCACAAAGAGUGGUUC CACGACAUCCCCCUGCCCUGGCAUGCCGGCGCUGAUACAGGCACACC CCACUGGAACAACAAAGAGGCUCUGGUGGAAUUCAAGGACGCCCAC GCCAAGCGGCAGACCGUGGUGGUGCUGGGAUCUCAGGAAGGCGCCG UGCAUACAGCUCUGGCUGGCGCCCUGGAAGCCGAAAUGGAUGGCGC CAAAGGCAGACUGUCCAGCGGCCACCUGAAGUGCCGGCUGAAGAUG GACAAGCUGCGGCUGAAGGGCGUGUCCUACAGCCUGUGUACCGCCGC CUUCACCUUCACCAAGAUCCCCGCCGAGACACUGCACGGCACCGUGA CUGUGGAAGUGCAGUACGCCGGCACCGACGGCCCUUGUAAAGUGCC UGCUCAGAUGGCCGUGGAUAUGCAGACCCUGACCCCCGUGGGCAGAC UGAUCACCGCCAACCCUGUGAUCACCGAGAGCACCGAGAACAGCAAG AUGAUGCUGGAACUGGACCCCCCCUUCGGCGACUCCUACAUCGUGAU CGGCGUGGGAGAGAAGAAGAUCACCCACCACUGGCACCGCAGCGGCA GCACAAUCGGCAAGGCCUUUGAAGCCACAGUGCGGGGAGCCAAGAG AAUGGCCGUGCUGGGAGAUACCGCCUGGGACUUUGGCUCUGUGGGC GGAGCCCUGAACUCUCUGGGCAAGGGAAUCCACCAGAUCUUCGGAG CCGCCUUUAAGAGCCUGUUCGGCGGCAUGAGCUGGUUCAGCCAGAU CCUGAUCGGCACCCUGCUCGUGUGGCUGGGCCUGAACACCAAGAACG GCAGCAUCUCCCUGACCUGCCUGGCUCUGGGAGGCGUGCUGAUCUUU CUGAGCACCGCCGUGUCUGCCUGAUAAUAGGCUGGAGCCUCGGUGG CCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCC UGCACCCGUACCCCCGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC HuIgG_(k) signal AUGGAAACCCCUGCCCAGCUGCUGUUCCUGCUGCUGCUGUGGCUGCC 148 peptide_prME #2 UGAUACCACCGGCACCAGAAGAGGCAGCGCCUACUACAUGUACCUGG (Brazil_isolate_ ACAGAAGCGACGCCGGCGAGGCCAUCAGCUUUCCAACCACCCUGGGC ZikaSPH2015), ORF AUGAACAAGUGCUACAUCCAGAUCAUGGACCUGGGCCACAUGUGCG Sequence, NT ACGCCACCAUGAGCUACGAGUGCCCCAUGCUGGACGAGGGCGUGGA ACCCGACGAUGUGGACUGCUGGUGCAACACCACCAGCACCUGGGUGG UGUACGGCACCUGUCACCACAAGAAGGGCGAAGCCAGACGGUCCAG ACGGGCCGUGACACUGCCUAGCCACUCCACCAGAAAGCUGCAGACCC GGUCCCAGACCUGGCUGGAAAGCAGAGAGUACACCAAGCACCUGAU CCGGGUGGAAAACUGGAUCUUCCGGAACCCCGGCUUUGCCCUGGCCG CUGCUGCUAUUGCUUGGCUGCUGGGCAGCAGCACCUCCCAGAAAGU GAUCUACCUCGUGAUGAUCCUGCUGAUCGCCCCUGCCUACAGCAUCC GGUGUAUCGGCGUGUCCAACCGGGACUUCGUGGAAGGCAUGAGCGG CGGCACAUGGGUGGACGUGGUGCUGGAACAUGGCGGCUGCGUGACA GUGAUGGCCCAGGAUAAGCCCGCCGUGGACAUCGAGCUCGUGACCAC CACCGUGUCCAAUAUGGCCGAAGUGCGGAGCUACUGCUACGAGGCC AGCAUCAGCGACAUGGCCAGCGACAGCAGAUGCCCUACACAGGGCGA GGCCUACCUGGAUAAGCAGUCCGACACCCAGUACGUGUGCAAGCGG ACCCUGGUGGAUAGAGGCUGGGGCAAUGGCUGCGGCCUGUUUGGCA AGGGCAGCCUCGUGACCUGCGCCAAGUUCGCCUGCAGCAAGAAGAU GACCGGCAAGAGCAUCCAGCCCGAGAACCUGGAAUACCGGAUCAUGC UGAGCGUGCACGGCAGCCAGCACUCCGGCAUGAUCGUGAACGACACC GGCCACGAGACAGACGAGAACCGGGCCAAGGUGGAAAUCACCCCCAA CAGCCCUAGAGCCGAGGCCACACUGGGCGGCUUUGGAUCUCUGGGCC UGGACUGCGAGCCUAGAACCGGCCUGGAUUUCAGCGACCUGUACUA CCUGACCAUGAACAACAAGCACUGGCUGGUGCACAAAGAGUGGUUC CACGACAUCCCCCUGCCCUGGCAUGCCGGCGCUGAUACAGGCACACC CCACUGGAACAACAAAGAGGCUCUGGUGGAAUUCAAGGACGCCCAC GCCAAGCGGCAGACCGUGGUGGUGCUGGGAUCUCAGGAAGGCGCCG UGCAUACAGCUCUGGCUGGCGCCCUGGAAGCCGAAAUGGAUGGCGC CAAAGGCAGACUGUCCAGCGGCCACCUGAAGUGCCGGCUGAAGAUG GACAAGCUGCGGCUGAAGGGCGUGUCCUACAGCCUGUGUACCGCCGC CUUCACCUUCACCAAGAUCCCCGCCGAGACACUGCACGGCACCGUGA CUGUGGAAGUGCAGUACGCCGGCACCGACGGCCCUUGUAAAGUGCC UGCUCAGAUGGCCGUGGAUAUGCAGACCCUGACCCCCGUGGGCAGAC UGAUCACCGCCAACCCUGUGAUCACCGAGAGCACCGAGAACAGCAAG AUGAUGCUGGAACUGGACCCCCCCUUCGGCGACUCCUACAUCGUGAU CGGCGUGGGAGAGAAGAAGAUCACCCACCACUGGCACCGCAGCGGCA GCACAAUCGGCAAGGCCUUUGAAGCCACAGUGCGGGGAGCCAAGAG AAUGGCCGUGCUGGGAGAUACCGCCUGGGACUUUGGCUCUGUGGGC GGAGCCCUGAACUCUCUGGGCAAGGGAAUCCACCAGAUCUUCGGAG CCGCCUUUAAGAGCCUGUUCGGCGGCAUGAGCUGGUUCAGCCAGAU CCUGAUCGGCACCCUGCUCGUGUGGCUGGGCCUGAACACCAAGAACG GCAGCAUCUCCCUGACCUGCCUGGCUCUGGGAGGCGUGCUGAUCUUU CUGAGCACCGCCGUGUCUGCC HuIgG_(k) signal G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCC 149 peptide_prME #2 ACCAUGGAAACCCCUGCCCAGCUGCUGUUCCUGCUGCUGCUGUGGCU (Brazil_isolate_ GCCUGAUACCACCGGCACCAGAAGAGGCAGCGCCUACUACAUGUACC ZikaSPH2015), mRNA UGGACAGAAGCGACGCCGGCGAGGCCAUCAGCUUUCCAACCACCCUG Sequence (T100 GGCAUGAACAAGUGCUACAUCCAGAUCAUGGACCUGGGCCACAUGU tail) GCGACGCCACCAUGAGCUACGAGUGCCCCAUGCUGGACGAGGGCGUG GAACCCGACGAUGUGGACUGCUGGUGCAACACCACCAGCACCUGGGU GGUGUACGGCACCUGUCACCACAAGAAGGGCGAAGCCAGACGGUCC AGACGGGCCGUGACACUGCCUAGCCACUCCACCAGAAAGCUGCAGAC CCGGUCCCAGACCUGGCUGGAAAGCAGAGAGUACACCAAGCACCUGA UCCGGGUGGAAAACUGGAUCUUCCGGAACCCCGGCUUUGCCCUGGCC GCUGCUGCUAUUGCUUGGCUGCUGGGCAGCAGCACCUCCCAGAAAG UGAUCUACCUCGUGAUGAUCCUGCUGAUCGCCCCUGCCUACAGCAUC CGGUGUAUCGGCGUGUCCAACCGGGACUUCGUGGAAGGCAUGAGCG GCGGCACAUGGGUGGACGUGGUGCUGGAACAUGGCGGCUGCGUGAC AGUGAUGGCCCAGGAUAAGCCCGCCGUGGACAUCGAGCUCGUGACC ACCACCGUGUCCAAUAUGGCCGAAGUGCGGAGCUACUGCUACGAGG CCAGCAUCAGCGACAUGGCCAGCGACAGCAGAUGCCCUACACAGGGC GAGGCCUACCUGGAUAAGCAGUCCGACACCCAGUACGUGUGCAAGC GGACCCUGGUGGAUAGAGGCUGGGGCAAUGGCUGCGGCCUGUUUGG CAAGGGCAGCCUCGUGACCUGCGCCAAGUUCGCCUGCAGCAAGAAGA UGACCGGCAAGAGCAUCCAGCCCGAGAACCUGGAAUACCGGAUCAU GCUGAGCGUGCACGGCAGCCAGCACUCCGGCAUGAUCGUGAACGACA CCGGCCACGAGACAGACGAGAACCGGGCCAAGGUGGAAAUCACCCCC AACAGCCCUAGAGCCGAGGCCACACUGGGCGGCUUUGGAUCUCUGG GCCUGGACUGCGAGCCUAGAACCGGCCUGGAUUUCAGCGACCUGUAC UACCUGACCAUGAACAACAAGCACUGGCUGGUGCACAAAGAGUGGU UCCACGACAUCCCCCUGCCCUGGCAUGCCGGCGCUGAUACAGGCACA CCCCACUGGAACAACAAAGAGGCUCUGGUGGAAUUCAAGGACGCCC ACGCCAAGCGGCAGACCGUGGUGGUGCUGGGAUCUCAGGAAGGCGC CGUGCAUACAGCUCUGGCUGGCGCCCUGGAAGCCGAAAUGGAUGGC GCCAAAGGCAGACUGUCCAGCGGCCACCUGAAGUGCCGGCUGAAGA UGGACAAGCUGCGGCUGAAGGGCGUGUCCUACAGCCUGUGUACCGC CGCCUUCACCUUCACCAAGAUCCCCGCCGAGACACUGCACGGCACCG UGACUGUGGAAGUGCAGUACGCCGGCACCGACGGCCCUUGUAAAGU GCCUGCUCAGAUGGCCGUGGAUAUGCAGACCCUGACCCCCGUGGGCA GACUGAUCACCGCCAACCCUGUGAUCACCGAGAGCACCGAGAACAGC AAGAUGAUGCUGGAACUGGACCCCCCCUUCGGCGACUCCUACAUCGU GAUCGGCGUGGGAGAGAAGAAGAUCACCCACCACUGGCACCGCAGC GGCAGCACAAUCGGCAAGGCCUUUGAAGCCACAGUGCGGGGAGCCA AGAGAAUGGCCGUGCUGGGAGAUACCGCCUGGGACUUUGGCUCUGU GGGCGGAGCCCUGAACUCUCUGGGCAAGGGAAUCCACCAGAUCUUC GGAGCCGCCUUUAAGAGCCUGUUCGGCGGCAUGAGCUGGUUCAGCC AGAUCCUGAUCGGCACCCUGCUCGUGUGGCUGGGCCUGAACACCAAG AACGGCAGCAUCUCCCUGACCUGCCUGGCUCUGGGAGGCGUGCUGAU CUUUCUGAGCACCGCCGUGUCUGCCUGAUAAUAGGCUGGAGCCUCG GUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCC CUUCCUGCACCCGUACCCCCGUGGUCUUUGAAUAAAGUCUGAGUGG GCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAUCUAG HuIgG_(k) signal UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAG 150 peptide_E GGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACC (Brazil_isolate_ AUGGAAACCCCUGCCCAGCUGCUGUUCCUGCUGCUGCUGUGGCUGCC ZikaSPH2015), UGACACCACCGGCAUCAGAUGCAUCGGCGUGUCCAACCGGGACUUCG Sequence, NT (5′ UGGAAGGCAUGAGCGGCGGCACAUGGGUGGACGUGGUGCUGGAACA UTR, ORF, 3′ UGGCGGCUGCGUGACAGUGAUGGCCCAGGAUAAGCCCGCCGUGGAC UTR) AUCGAGCUCGUGACCACCACCGUGUCCAAUAUGGCCGAAGUGCGGA GCUACUGCUACGAGGCCAGCAUCAGCGACAUGGCCAGCGACAGCAGA UGCCCUACACAGGGCGAGGCCUACCUGGACAAGCAGAGCGACACCCA GUACGUGUGCAAGCGGACCCUGGUGGAUAGAGGCUGGGGCAAUGGC UGCGGCCUGUUUGGCAAGGGCAGCCUCGUGACCUGCGCCAAGUUCGC CUGCAGCAAGAAGAUGACCGGCAAGAGCAUCCAGCCCGAGAACCUG GAAUACCGGAUCAUGCUGAGCGUGCACGGCAGCCAGCACUCCGGCAU GAUCGUGAACGACACCGGCCACGAGACAGACGAGAACCGGGCCAAG GUGGAAAUCACCCCCAACAGCCCUAGAGCCGAGGCCACACUGGGCGG CUUUGGAUCUCUGGGCCUGGACUGCGAGCCUAGAACCGGCCUGGAU UUCAGCGACCUGUACUACCUGACCAUGAACAACAAGCACUGGCUGG UGCACAAAGAGUGGUUCCACGACAUCCCCCUGCCCUGGCAUGCCGGC GCUGAUACAGGCACACCCCACUGGAACAACAAAGAGGCUCUGGUGG AAUUCAAGGACGCCCACGCCAAGCGGCAGACCGUGGUGGUGCUGGG AUCUCAGGAAGGCGCCGUGCAUACAGCUCUGGCUGGCGCCCUGGAA GCCGAAAUGGAUGGCGCCAAAGGCAGACUGAGCAGCGGCCACCUGA AGUGCCGGCUGAAGAUGGACAAGCUGCGGCUGAAGGGCGUGUCCUA CAGCCUGUGUACCGCCGCCUUCACCUUCACCAAGAUCCCCGCCGAGA CACUGCACGGCACCGUGACUGUGGAAGUGCAGUACGCCGGCACCGAC GGCCCUUGUAAAGUGCCUGCUCAGAUGGCCGUGGAUAUGCAGACCC UGACCCCCGUGGGCAGGCUGAUCACAGCCAACCCUGUGAUCACCGAG AGCACCGAGAACAGCAAGAUGAUGCUGGAACUGGACCCCCCCUUCGG CGACUCCUACAUCGUGAUCGGCGUGGGAGAGAAGAAGAUCACCCAC CACUGGCACAGAAGCGGCAGCACCAUCGGCAAGGCCUUUGAGGCUAC AGUGCGGGGAGCCAAGAGAAUGGCCGUGCUGGGAGAUACCGCCUGG GACUUUGGCUCUGUGGGCGGAGCCCUGAACUCUCUGGGCAAGGGAA UCCACCAGAUCUUCGGCGCUGCCUUCAAGAGCCUGUUCGGCGGCAUG AGCUGGUUCAGCCAGAUCCUGAUCGGCACCCUGCUCGUGUGGCUGG GCCUGAACACCAAGAACGGCAGCAUCUCCCUGACCUGCCUGGCUCUG GGAGGCGUGCUGAUCUUUCUGAGCACCGCCGUGUCUGCCUGAUAAU AGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCC CAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCGUGGUCUUUGAAU AAAGUCUGAGUGGGCGGC HuIgG_(k) signal AUGGAAACCCCUGCCCAGCUGCUGUUCCUGCUGCUGCUGUGGCUGCC 151 peptide_E UGACACCACCGGCAUCAGAUGCAUCGGCGUGUCCAACCGGGACUUCG (Brazil_isolate_ UGGAAGGCAUGAGCGGCGGCACAUGGGUGGACGUGGUGCUGGAACA ZikaSPH2015), ORF UGGCGGCUGCGUGACAGUGAUGGCCCAGGAUAAGCCCGCCGUGGAC Sequence, NT AUCGAGCUCGUGACCACCACCGUGUCCAAUAUGGCCGAAGUGCGGA GCUACUGCUACGAGGCCAGCAUCAGCGACAUGGCCAGCGACAGCAGA UGCCCUACACAGGGCGAGGCCUACCUGGACAAGCAGAGCGACACCCA GUACGUGUGCAAGCGGACCCUGGUGGAUAGAGGCUGGGGCAAUGGC UGCGGCCUGUUUGGCAAGGGCAGCCUCGUGACCUGCGCCAAGUUCGC CUGCAGCAAGAAGAUGACCGGCAAGAGCAUCCAGCCCGAGAACCUG GAAUACCGGAUCAUGCUGAGCGUGCACGGCAGCCAGCACUCCGGCAU GAUCGUGAACGACACCGGCCACGAGACAGACGAGAACCGGGCCAAG GUGGAAAUCACCCCCAACAGCCCUAGAGCCGAGGCCACACUGGGCGG CUUUGGAUCUCUGGGCCUGGACUGCGAGCCUAGAACCGGCCUGGAU UUCAGCGACCUGUACUACCUGACCAUGAACAACAAGCACUGGCUGG UGCACAAAGAGUGGUUCCACGACAUCCCCCUGCCCUGGCAUGCCGGC GCUGAUACAGGCACACCCCACUGGAACAACAAAGAGGCUCUGGUGG AAUUCAAGGACGCCCACGCCAAGCGGCAGACCGUGGUGGUGCUGGG AUCUCAGGAAGGCGCCGUGCAUACAGCUCUGGCUGGCGCCCUGGAA GCCGAAAUGGAUGGCGCCAAAGGCAGACUGAGCAGCGGCCACCUGA AGUGCCGGCUGAAGAUGGACAAGCUGCGGCUGAAGGGCGUGUCCUA CAGCCUGUGUACCGCCGCCUUCACCUUCACCAAGAUCCCCGCCGAGA CACUGCACGGCACCGUGACUGUGGAAGUGCAGUACGCCGGCACCGAC GGCCCUUGUAAAGUGCCUGCUCAGAUGGCCGUGGAUAUGCAGACCC UGACCCCCGUGGGCAGGCUGAUCACAGCCAACCCUGUGAUCACCGAG AGCACCGAGAACAGCAAGAUGAUGCUGGAACUGGACCCCCCCUUCGG CGACUCCUACAUCGUGAUCGGCGUGGGAGAGAAGAAGAUCACCCAC CACUGGCACAGAAGCGGCAGCACCAUCGGCAAGGCCUUUGAGGCUAC AGUGCGGGGAGCCAAGAGAAUGGCCGUGCUGGGAGAUACCGCCUGG GACUUUGGCUCUGUGGGCGGAGCCCUGAACUCUCUGGGCAAGGGAA UCCACCAGAUCUUCGGCGCUGCCUUCAAGAGCCUGUUCGGCGGCAUG AGCUGGUUCAGCCAGAUCCUGAUCGGCACCCUGCUCGUGUGGCUGG GCCUGAACACCAAGAACGGCAGCAUCUCCCUGACCUGCCUGGCUCUG GGAGGCGUGCUGAUCUUUCUGAGCACCGCCGUGUCUGCC HuIgG_(k) signal G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCC 152 peptide_E ACCAUGGAAACCCCUGCCCAGCUGCUGUUCCUGCUGCUGCUGUGGCU (Brazil_isolate_ GCCUGACACCACCGGCAUCAGAUGCAUCGGCGUGUCCAACCGGGACU ZikaSPH2015), mRNA UCGUGGAAGGCAUGAGCGGCGGCACAUGGGUGGACGUGGUGCUGGA Sequence (T100 ACAUGGCGGCUGCGUGACAGUGAUGGCCCAGGAUAAGCCCGCCGUG tail) GACAUCGAGCUCGUGACCACCACCGUGUCCAAUAUGGCCGAAGUGCG GAGCUACUGCUACGAGGCCAGCAUCAGCGACAUGGCCAGCGACAGCA GAUGCCCUACACAGGGCGAGGCCUACCUGGACAAGCAGAGCGACACC CAGUACGUGUGCAAGCGGACCCUGGUGGAUAGAGGCUGGGGCAAUG GCUGCGGCCUGUUUGGCAAGGGCAGCCUCGUGACCUGCGCCAAGUUC GCCUGCAGCAAGAAGAUGACCGGCAAGAGCAUCCAGCCCGAGAACCU GGAAUACCGGAUCAUGCUGAGCGUGCACGGCAGCCAGCACUCCGGCA UGAUCGUGAACGACACCGGCCACGAGACAGACGAGAACCGGGCCAA GGUGGAAAUCACCCCCAACAGCCCUAGAGCCGAGGCCACACUGGGCG GCUUUGGAUCUCUGGGCCUGGACUGCGAGCCUAGAACCGGCCUGGA UUUCAGCGACCUGUACUACCUGACCAUGAACAACAAGCACUGGCUG GUGCACAAAGAGUGGUUCCACGACAUCCCCCUGCCCUGGCAUGCCGG CGCUGAUACAGGCACACCCCACUGGAACAACAAAGAGGCUCUGGUG GAAUUCAAGGACGCCCACGCCAAGCGGCAGACCGUGGUGGUGCUGG GAUCUCAGGAAGGCGCCGUGCAUACAGCUCUGGCUGGCGCCCUGGA AGCCGAAAUGGAUGGCGCCAAAGGCAGACUGAGCAGCGGCCACCUG AAGUGCCGGCUGAAGAUGGACAAGCUGCGGCUGAAGGGCGUGUCCU ACAGCCUGUGUACCGCCGCCUUCACCUUCACCAAGAUCCCCGCCGAG ACACUGCACGGCACCGUGACUGUGGAAGUGCAGUACGCCGGCACCGA CGGCCCUUGUAAAGUGCCUGCUCAGAUGGCCGUGGAUAUGCAGACC CUGACCCCCGUGGGCAGGCUGAUCACAGCCAACCCUGUGAUCACCGA GAGCACCGAGAACAGCAAGAUGAUGCUGGAACUGGACCCCCCCUUCG GCGACUCCUACAUCGUGAUCGGCGUGGGAGAGAAGAAGAUCACCCA CCACUGGCACAGAAGCGGCAGCACCAUCGGCAAGGCCUUUGAGGCUA CAGUGCGGGGAGCCAAGAGAAUGGCCGUGCUGGGAGAUACCGCCUG GGACUUUGGCUCUGUGGGCGGAGCCCUGAACUCUCUGGGCAAGGGA AUCCACCAGAUCUUCGGCGCUGCCUUCAAGAGCCUGUUCGGCGGCAU GAGCUGGUUCAGCCAGAUCCUGAUCGGCACCCUGCUCGUGUGGCUG GGCCUGAACACCAAGAACGGCAGCAUCUCCCUGACCUGCCUGGCUCU GGGAGGCGUGCUGAUCUUUCUGAGCACCGCCGUGUCUGCCUGAUAA UAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCC CCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCGUGGUCUUUGAA UAAAGUCUGAGUGGGCGGCAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAUCUAG Zika_RIO- AUGCUGGGCAGCAACAGCGGCCAGAGAGUGGUGUUCACCAUCCUGC 153 U1_JEVsp UGCUGCUGGUGGCCCCUGCCUACAGCGCCGAAGUGACAAGAAGAGG Zika PRME Strain CAGCGCCUACUACAUGUACCUGGACCGGAACGAUGCCGGCGAGGCCA ascension id: UCAGCUUUCCAACCACCCUGGGCAUGAACAAGUGCUACAUCCAGAUC ANG09399 with AUGGACCUGGGCCACAUGUGCGACGCCACCAUGAGCUACGAGUGCCC JEV PRM signal CAUGCUGGACGAGGGCGUGGAACCCGACGAUGUGGACUGCUGGUGC sequence AAUACCACCAGCACCUGGGUGGUGUACGGCACCUGUCACCACAAGAA (optimized) GGGCGAAGCCAGACGGUCCAGACGGGCCGUGACACUGCCUAGCCACA GCACCAGAAAGCUGCAGACCCGGUCCCAGACCUGGCUGGAAAGCAGA GAGUACACCAAGCACCUGAUCCGGGUGGAAAACUGGAUCUUCCGGA ACCCCGGCUUUGCCCUGGCUGCCGCUGCUAUUGCUUGGCUGCUGGGC UCUAGCACCAGCCAGAAAGUGAUCUACCUCGUGAUGAUCCUGCUGA UCGCCCCAGCCUACUCCAUCCGGUGUAUCGGCGUGUCCAACCGGGAC UUCGUGGAAGGCAUGAGCGGCGGCACAUGGGUGGACGUGGUGCUGG AACAUGGCGGCUGCGUGACAGUGAUGGCCCAGGACAAGCCCACCGU GGACAUCGAGCUCGUGACCACCACCGUGUCCAAUAUGGCCGAAGUGC GGAGCUACUGCUACGAGGCCAGCAUCAGCGACAUGGCCAGCGACAGC AGAUGCCCUACACAGGGCGAGGCCUACCUGGACAAGCAGUCCGACAC CCAGUACGUGUGCAAGCGGACCCUGGUGGACAGGGGCUGGGGCAAU GGCUGUGGCCUGUUUGGCAAGGGCAGCCUCGUGACCUGCGCCAAGU UCGCCUGCAGCAAGAAGAUGACCGGCAAGAGCAUCCAGCCCGAGAAC CUGGAAUACCGGAUCAUGCUGAGCGUGCACGGCUCCCAGCACAGCGG CAUGAUCGUGAACGACACCGGCCACGAGACAGACGAGAACCGGGCCA AGGUGGAAAUCACCCCCAACAGCCCUAGAGCCGAGGCCACACUGGGC GGCUUUGGAUCUCUGGGCCUGGACUGCGAGCCUAGAACCGGCCUGG AUUUCAGCGACCUGUACUACCUGACCAUGAACAACAAACACUGGCU GGUGCACAAAGAGUGGUUCCACGACAUCCCCCUGCCCUGGCAUGCUG GCGCUGAUACAGGCACCCCCCACUGGAACAACAAAGAGGCCCUGGUG GAAUUCAAGGACGCCCACGCCAAGCGGCAGACCGUGGUGGUGCUGG GAUCUCAGGAAGGCGCCGUGCAUACAGCUCUGGCUGGCGCCCUGGA AGCCGAAAUGGAUGGCGCCAAAGGCAGACUGAGCAGCGGCCACCUG AAGUGCCGGCUGAAGAUGGACAAGCUGCGGCUGAAGGGCGUGUCCU ACAGCCUGUGUACCGCCGCCUUCACCUUCACCAAGAUCCCCGCCGAG ACACUGCACGGCACCGUGACUGUGGAAGUGCAGUACGCCGGCACCGA CGGCCCUUGUAAAGUGCCUGCUCAGAUGGCCGUGGAUAUGCAGACC CUGACCCCCGUGGGCAGGCUGAUCACAGCCAACCCUGUGAUCACCGA GAGCACCGAGAACAGCAAGAUGAUGCUGGAACUGGACCCCCCCUUCG GCGACUCCUACAUCGUGAUCGGCGUGGGAGAGAAGAAGAUCACCCA CCACUGGCACAGAAGCGGCAGCACCAUCGGCAAGGCCUUUGAGGCUA CAGUGCGGGGAGCCAAGAGAAUGGCCGUGCUGGGCGAUACCGCCUG GGAUUUUGGCUCUGUGGGCGGAGCCCUGAACAGCCUGGGAAAGGGC AUCCACCAGAUCUUCGGAGCCGCCUUUAAGAGCCUGUUCGGCGGCAU GAGCUGGUUCAGCCAGAUCCUGAUCGGCACCCUGCUGAUGUGGCUG GGCCUGAACACCAAGAACGGCAGCAUCUCCCUGAUGUGCCUGGCUCU GGGCGGCGUGCUGAUCUUUCUGAGCACAGCCGUGUCCGCC Zika_RIO- AUGAAGUGCCUGCUGUACCUGGCCUUCCUGUUCAUCGGCGUGAACU 154 U1_VSVgSp GCGCCGAAGUGACCAGAAGAGGCAGCGCCUACUACAUGUACCUGGA Zika PRME Strain CCGGAACGAUGCCGGCGAGGCCAUCAGCUUUCCAACCACCCUGGGCA ascension id: UGAACAAGUGCUACAUCCAGAUCAUGGACCUGGGCCACAUGUGCGA ANG09399 with CGCCACCAUGAGCUACGAGUGCCCCAUGCUGGACGAGGGCGUGGAAC VSVg protein CCGACGAUGUGGACUGCUGGUGCAACACCACCAGCACCUGGGUGGU signal sequence GUACGGCACCUGUCACCACAAGAAGGGCGAAGCCAGACGGUCCAGAC (optimized) GGGCCGUGACACUGCCUAGCCACAGCACCAGAAAGCUGCAGACCCGG UCCCAGACCUGGCUGGAAAGCAGAGAGUACACCAAGCACCUGAUCCG GGUGGAAAACUGGAUCUUCCGGAACCCCGGCUUUGCCCUGGCCGCUG CUGCUAUUGCUUGGCUGCUGGGCAGCAGCACCUCCCAGAAAGUGAU CUACCUCGUGAUGAUCCUGCUGAUCGCCCCUGCCUACAGCAUCCGGU GUAUCGGCGUGUCCAACCGGGACUUCGUGGAAGGCAUGAGCGGCGG CACAUGGGUGGACGUGGUGCUGGAACAUGGCGGCUGCGUGACAGUG AUGGCCCAGGACAAGCCCACCGUGGACAUCGAGCUCGUGACCACCAC CGUGUCCAAUAUGGCCGAAGUGCGGAGCUACUGCUACGAGGCCAGC AUCAGCGACAUGGCCAGCGACAGCAGAUGCCCUACACAGGGCGAGGC CUACCUGGACAAGCAGUCCGACACCCAGUACGUGUGCAAGCGGACCC UGGUGGAUAGAGGCUGGGGCAAUGGCUGCGGCCUGUUUGGCAAGGG CAGCCUCGUGACCUGUGCCAAGUUCGCCUGCAGCAAGAAGAUGACCG GCAAGAGCAUCCAGCCCGAGAACCUGGAAUACCGGAUCAUGCUGAG CGUGCACGGCAGCCAGCACUCCGGCAUGAUCGUGAACGACACCGGCC ACGAGACAGACGAGAACCGGGCCAAGGUGGAAAUCACCCCCAACAGC CCUAGAGCCGAGGCCACACUGGGCGGCUUUGGAUCUCUGGGCCUGG ACUGCGAGCCUAGAACCGGCCUGGAUUUCAGCGACCUGUACUACCUG ACCAUGAACAACAAGCACUGGCUGGUGCACAAAGAGUGGUUCCACG ACAUCCCCCUGCCCUGGCAUGCCGGCGCUGAUACAGGCACACCCCAC UGGAACAACAAAGAGGCUCUGGUGGAAUUCAAGGACGCCCACGCCA AGCGGCAGACCGUGGUGGUGCUGGGAUCUCAGGAAGGCGCCGUGCA UACAGCUCUGGCUGGCGCCCUGGAAGCCGAAAUGGAUGGCGCCAAA GGCAGACUGUCCAGCGGCCACCUGAAGUGCAGACUGAAGAUGGACA AGCUGCGGCUGAAGGGCGUGUCCUACAGCCUGUGUACCGCCGCCUUC ACCUUCACCAAGAUCCCCGCCGAGACACUGCACGGCACCGUGACUGU GGAAGUGCAGUACGCCGGCACCGACGGCCCUUGUAAAGUGCCAGCUC AGAUGGCCGUGGAUAUGCAGACCCUGACCCCCGUGGGCAGACUGAU CACCGCCAACCCUGUGAUCACCGAGAGCACCGAGAACAGCAAGAUGA UGCUGGAACUGGACCCCCCCUUCGGCGACUCCUACAUCGUGAUCGGC GUGGGAGAGAAGAAGAUCACCCACCACUGGCACAGAAGCGGCAGCA CCAUCGGCAAGGCCUUUGAGGCUACAGUGCGGGGAGCCAAGAGAAU GGCCGUGCUGGGAGAUACCGCCUGGGACUUUGGCUCUGUGGGCGGA GCCCUGAACUCUCUGGGCAAGGGAAUCCACCAGAUCUUCGGAGCCGC CUUUAAGAGCCUGUUCGGCGGCAUGAGCUGGUUCAGCCAGAUCCUG AUCGGCACCCUGCUGAUGUGGCUGGGCCUGAACACCAAGAACGGCA GCAUCUCCCUGAUGUGCCUGGCUCUGGGAGGCGUGCUGAUCUUCCU GAGCACAGCCGUGUCUGCC ZIKA_PRME_DSP_ AUGGACUGGACCUGGAUCCUGUUCCUGGUGGCCGCUGCCACAAGAG 155 N154A UGCACAGCGUGGAAGUGACCAGACGGGGCAGCGCCUACUACAUGUA Zika PRME Strain CCUGGACAGAAGCGACGCCGGCGAGGCCAUCAGCUUUCCAACCACCC ascension id: UGGGCAUGAACAAGUGCUACAUCCAGAUCAUGGACCUGGGCCACAU ACD75819 with GUGCGACGCCACCAUGAGCUACGAGUGCCCCAUGCUGGACGAGGGCG IgE signal peptide UGGAACCCGACGAUGUGGACUGCUGGUGCAACACCACCAGCACCUGG (optimized) GUGGUGUACGGCACCUGUCACCACAAGAAGGGCGAAGCCAGACGGU CCAGACGGGCCGUGACACUGCCUAGCCACAGCACCAGAAAGCUGCAG ACCCGGUCCCAGACCUGGCUGGAAAGCAGAGAGUACACCAAGCACCU GAUCCGGGUGGAAAACUGGAUCUUCCGGAACCCCGGCUUUGCCCUG GCUGCCGCUGCUAUUGCUUGGCUGCUGGGCAGCAGCACCUCCCAGAA AGUGAUCUACCUCGUGAUGAUCCUGCUGAUCGCCCCUGCCUACAGCA UCCGGUGUAUCGGCGUGUCCAACCGGGACUUCGUGGAAGGCAUGAG CGGCGGCACAUGGGUGGACGUGGUGCUGGAACAUGGCGGCUGCGUG ACAGUGAUGGCCCAGGAUAAGCCCGCCGUGGACAUCGAGCUCGUGA CCACCACCGUGUCCAAUAUGGCCGAAGUGCGGAGCUACUGCUACGAG GCCAGCAUCAGCGACAUGGCCAGCGACAGCAGAUGCCCUACACAGGG CGAGGCCUACCUGGAUAAGCAGUCCGACACCCAGUACGUGUGCAAGC GGACCCUGGUGGAUAGAGGCUGGGGCAAUGGCUGCGGCCUGUUUGG CAAGGGCAGCCUCGUGACCUGCGCCAAGUUCGCCUGCAGCAAGAAGA UGACCGGCAAGAGCAUCCAGCCCGAGAACCUGGAAUACCGGAUCAU GCUGAGCGUGCACGGCUCCCAGCACAGCGGCAUGAUCGUGGCCGACA CCGGCCACGAGACAGACGAGAACCGGGCCAAGGUGGAAAUCACCCCC AACAGCCCUAGAGCCGAGGCCACACUGGGCGGCUUUGGAUCUCUGG GCCUGGACUGCGAGCCUAGAACCGGCCUGGAUUUCAGCGACCUGUAC UACCUGACCAUGAACAACAAGCACUGGCUGGUGCACAAAGAGUGGU UCCACGACAUCCCCCUGCCCUGGCAUGCUGGCGCUGAUACAGGCACC CCCCACUGGAACAACAAAGAGGCUCUGGUGGAAUUCAAGGACGCCC ACGCCAAGCGGCAGACCGUGGUGGUGCUGGGAUCUCAGGAAGGCGC CGUGCAUACAGCUCUGGCUGGCGCCCUGGAAGCCGAAAUGGAUGGC GCCAAAGGCAGACUGUCCUCCGGCCACCUGAAGUGCCGGCUGAAGAU GGACAAGCUGCGGCUGAAGGGCGUGUCCUACAGCCUGUGUACCGCC GCCUUCACCUUCACCAAGAUCCCCGCCGAGACACUGCACGGCACCGU GACUGUGGAAGUGCAGUACGCCGGCACCGACGGCCCUUGUAAAGUG CCUGCUCAGAUGGCCGUGGAUAUGCAGACCCUGACCCCCGUGGGCAG ACUGAUCACCGCCAACCCUGUGAUCACCGAGAGCACCGAGAACAGCA AGAUGAUGCUGGAACUGGACCCCCCCUUCGGCGACUCCUACAUCGUG AUCGGCGUGGGAGAGAAGAAGAUCACCCACCACUGGCACAGAUCCG GCAGCACCAUCGGCAAGGCCUUUGAGGCUACAGUGCGGGGAGCCAA GAGAAUGGCCGUGCUGGGCGAUACCGCCUGGGAUUUUGGCUCUGUG GGCGGAGCCCUGAACAGCCUGGGAAAGGGCAUCCACCAGAUCUUCG GCGCUGCCUUCAAGAGCCUGUUCGGCGGCAUGAGCUGGUUCAGCCA GAUCCUGAUCGGCACCCUGCUCGUGUGGCUGGGCCUGAACACCAAGA ACGGCAGCAUCUCCCUGACCUGCCUGGCUCUGGGCGGCGUGCUGAUC UUUCUGAGCACAGCCGUGUCCGCC

TABLE 26 ZIKV Amino Acid Sequences SEQ ID Description Sequence NO: FSM|ACD75819 MKNPKEEIRRIRIVNMLKRGVARVSPFGGLKRLPAGLLLGHGPIRM 156 polyprotein VLAILAFLRFTAIKPSLGLINRWGSVGKKEAMEIIKKFKKDLAAMLR IINARKEKKRRGTDTSVGIVGLLLTTAMAVEVTRRGSAYYMYLDRS DAGEAISFPTTLGMNKCYIQIMDLGHMCDATMSYECPMLDEGVEP DDVDCWCNTTSTWVVYGTCHHKKGEARRSRRAVTLPSHSTRKLQ TRSQTWLESREYTKHLIRVENWIFRNPGFALAAAAIAWLLGSSTSQ KVIYLVMILLIAPAYSIRCIGVSNRDFVEGMSGGTWVDVVLEHGGC VTVMAQDKPAVDIELVTTTVSNMAEVRSYCYEASISDMASDSRCPT QGEAYLDKQSDTQYVCKRTLVDRGWGNGCGLFGKGSLVTCAKFA CSKKMTGKSIQPENLEYRIMLSVHGSQHSGMIVNDTGHETDENRAK VEITPNSPRAEATLGGFGSLGLDCEPRTGLDFSDLYYLTMNNKHWL VHKEWFHDIPLPWHAGADTGTPHWNNKEALVEFKDAHAKRQTVV VLGSQEGAVHTALAGALEAEMDGAKGRLSSGHLKCRLKMDKLRL KGVSYSLCTAAFTFTKIPAETLHGTVTVEVQYAGTDGPCKVPAQM AVDMQTLTPVGRLITANPVITESTENSKMMLELDPPFGDSYIVIGVG EKKITHHWHRSGSTIGKAFEATVRGAKRMAVLGDTAWDFGSVGG ALNSLGKGIHQIFGAAFKSLFGGMSWFSQILIGTLLVWLGLNTKNGS ISLTCLALGGVLIFLSTAVSA MR_766|ABI54475 MKNPKKKSGGFRIVNMLKRGVARVNPLGGLKRLPAGLLLGHGPIR 157 MVLAILAFLRFTAIKPSLGLINRWGTVGKKEAMEIIKKFKKDLAAM LRIINARKERKRRGADTSIGIVGLLLTTAMAAEITRRGSAYYMYLDR SDAGKAISFATTLGVNKCHVQIMDLGHMCDATMSYECPMLDEGVE PDDVDCWCNTTSTWVVYGTCHHKKGEARRSRRAVTLPSHSTRKL QTRSQTWLESREYTKHLIKVENWIFRNPGFTLVAVAIAWLLGSSTS QKVIYLVMILLIAPAYSIRCIGVSNRDFVEGMSGGTWVDVVLEHGG CVTVMAQDKPTVDIELVTTTVSNMAEVRSYCYEASISDMASDSRCP TQGEAYLDKQSDTQYVCKRTLVDRGWGNGCGLFGKGSLVTCAKF TCSKKMTGKSIQPENLEYRIMLSVHGSQHSGMIVNDENRAKVEVTP NSPRAEATLGGFGSLGLDCEPRTGLDFSDLYYLTMNNKHWLVHKE WFHDIPLPWHAGADTGTPHWNNKEALVEFKDAHAKRQTVVVLGS QEGAVHTALAGALEAEMDGAKGRLFSGHLKCRLKMDKLRLKGVS YSLCTAAFTFTKVPAETLHGTVTVEVQYAGTDGPCKVPAQMAVD MQTLTPVGRLITANPVITESTENSKMMLELDPPFGDSYIVIGVGDKK ITHHWHRSGSTIGKAFEATVRGAKRMAVLGDTAWDFGSVGGVFNS LGKGIHQIFGAAFKSLFGGMSWFSQILIGTLLVWLGLNTKNGSISLT CLALGGVMIFLSTAVSA SM_6_V1|ABI54480 MKNPKRAGSSRLVNMLRRGAARVIPPGGGLKRLPVGLLLGRGPIK 158 MILAILAFLRFTAIKPSTGLINRWGKVGKKEAIKILTKFKADVGTML RIINNRKTKKRGVETGIVFLALLVSIVAVEVTKKGDTYYMFADKKD AGKVVTFETESGPNRCSIQAMDIGHMCPATMSYECPVLEPQYEPED VDCWCNSTAAWIVYGTCTHKTTGETRRSRRSITLPSHASQKLETRS STWLESREYSKYLIKVENWILRNPGYALVAAVIGWTLGSSRSQKIIF VTLLMLVAPAYSIRCIGIGNRDFIEGMSGGTWVDIVLEHGGCVTVM SNDKPTLDFELVTTTASNMAEVRSYCYEANISEMASDSRCPTQGEA YLDKMADSQFVCKRGYVDRGWGNGCGLFGKGSIVTCAKFTCVKK LTGKSIQPENLEYRVLVSVHASQHGGMINNDTNHQHDKENRARIDI TASAPRVEVELGSFGSFSMECEPRSGLNFGDLYYLTMNNKHWLVN RDWFHDLSLPWHTGATSNNHHWNNKEALVEFREAHAKKQTAVVL GSQEGAVHAALAGALEAESDGHKATIYSGHLKCRLKLDKLRLKGM SYALCTGAFTFARTPSETIHGTATVELQYAGEDGPCKVPIVITSDTNS MASTGRLITANPVVTESGANSKMMVEIDPPFGDSYIIVGTGTTKITH HWHRAGSSIGRAFEATMRGAKRMAVLGDTAWDFGSVGGMFNSV GKFVHQVFGSAFKALFGGMSWFTQLLIGFLLIWMGLNARGGTVAM SFMGIGAMLIFLATSVSG MR_766|AAV34151 MKNPKEEIRRIRIVNMLKRGVARVNPLGGLKRLPAGLLLGHGPIRM 159 VLAILAFLRFTAIKPSLGLINRWGSVGKKEAMEIIKKFKKDLAAMLR IINARKERKRRGADTSIGIIGLLLTTAMAAEITRRGSAYYMYLDRSD AGKAISFATTLGVNKCHVQIMDLGHMCDATMSYECPMLDEGVEPD DVDCWCNTTSTWVVYGTCHHKKGEARRSRRAVTLPSHSTRKLQT RSQTWLESREYTKHLIKVENWIFRNPGFALVAVAIAWLLGSSTSQK VIYLVMILLIAPAYSIRCIGVSNRDFVEGMSGGTWVDVVLEHGGCV TVMAQDKPTVDIELVTTTVSNMAEVRSYCYEASISDMASDSRCPTQ GEAYLDKQSDTQYVCKRTLVDRGWGNGCGLFGKGSLVTCAKFTC SKKMTGKSIQPENLEYRIMLSVHGSQHSGMIGYETDEDRAKVEVTP NSPRAEATLGGFGSLGLDCEPRTGLDFSDLYYLTMNNKHWLVHKE WFHDIPLPWHAGADTGTPHWNNKEALVEFKDAHAKRQTVVVLGS QEGAVHTALAGALEAEMDGAKGRLFSGHLKCRLKMDKLRLKGVS YSLCTAAFTFTKVPAETLHGTVTVEVQYAGTDGPCKIPVQMAVDM QTLTPVGRLITANPVITESTENSKMMLELDPPFGDSYIVIGVGDKKIT HHWHRSGSTIGKAFEATVRGAKRMAVLGDTAWDFGSVGGVFNSL GKGIHQIFGAAFKSLFGGMSWFSQILIGTLLVWLGLNTKNGSISLTC LALGGVMIFLSTAVSA MR_766|YP_002790881 MKNPKEEIRRIRIVNMLKRGVARVNPLGGLKRLPAGLLLGHGPIRM 160 VLAILAFLRFTAIKPSLGLINRWGSVGKKEAMEIIKKFKKDLAAMLR IINARKERKRRGADTSIGIIGLLLTTAMAAEITRRGSAYYMYLDRSD AGKAISFATTLGVNKCHVQIMDLGHMCDATMSYECPMLDEGVEPD DVDCWCNTTSTWVVYGTCHHKKGEARRSRRAVTLPSHSTRKLQT RSQTWLESREYTKHLIKVENWIFRNPGFALVAVAIAWLLGSSTSQK VIYLVMILLIAPAYSIRCIGVSNRDFVEGMSGGTWVDVVLEHGGCV TVMAQDKPTVDIELVTTTVSNMAEVRSYCYEASISDMASDSRCPTQ GEAYLDKQSDTQYVCKRTLVDRGWGNGCGLFGKGSLVTCAKFTC KKMTGKSIQPENLEYRIMLSVHGSQHSGMIGYETDEDRAKVEVTPN SPRAEATLGGFGSLGLDCEPRTGLDFSDLYYLTMNNKHWLVHKEW FHDIPLPWHAGADTGTPHWNNKEALVEFKDAHAKRQTVVVLGSQ EGAVHTALAGALEAEMDGAKGRLFSGHLKCRLKMDKLRLKGVSY SLCTAAFTFTKVPAETLHGTVTVEVQYAGTDGPCKIPVQMAVDMQ TLTPVGRLITANPVITESTENSKMMLELDPPFGDSYIVIGVGDKKITH HWHRSGSTIGKAFEATVRGAKRMAVLGDTAWDFGSVGGVFNSLG KGIHQIFGAAFKSLFGGMSWFSQILIGTLLVWLGLNTKNGSISLTCL ALGGVMIFLSTAVSA ARB7701|AHF49785 MKNPKKKSGGFRIVNMLKRGVARVNPLGGLKRLPAGLLLGHGPIR 161 MVLAILAFLRFTAIKPSLGLINRWGSVGKKEAMEIIKKFKKDLAAM LRIINARKERKRRGADTSIGIIGLLLTTAMAAEITRRGSAYYMYLDR SDAGKAISFATNLGVNKCHVQIMDLGHMCDATMSYECPMLDEGV EPDDVDCWCNTTSTWVVYGTCHHKKGEARRSRRAVTLPSHSTRKL QTRSQTWLESREYTKHLIKVENWIFRNPGFALAAVAIAWLLGSSTS QKVIYLVMILLIAPAYSIRCIGVSNRDFVEGMSGGTWVDVVLEHGG CVTVMAQDKPTVDIELVTTTVSNMAEVRSYCYEASISDMASDSRCP TQGEAYLDKQSDTQYVCKRTLVDRGWGNGCGLFGKGSLVTCAKF TCSKKMTGKSIQPENLEYRIMLSVHGSQHSGMIVNDIGHETDENRA KVEVTPNSPRAEATLGGFGSLGLDCEPRTGLDFSDLYYLTMNNKH WLVHKEWFHDIPLPWHAGADTGTPHWNNKEALVEFKDAHAKRQT VVVLGSQEGAVHTALAGALEAEMDGAKGRLFSGHLKCRLKMDKL RLKGVSYSLCTAAFTFTKVPAETLHGTVTVEVQYAGTDGPCKVPA QMAVDMQTLTPVGRLITANPVITESTENSKMMLELDPPFGDSYIVIG VGDKKITHHWHRSGSTIGKAFEATVRGAKRMAVLGDTAWDFGSV GGVFNSLGKGVHQIFGAAFKSLFGGMSWFSQILIGTLLVWLGLNTK NGSISLTCLALGGVMIFLSTAVSA ARB15076|AHF49784 MKNPKKKSGGFRIVNMLKRGVARVNPLGGLKRLPAGLLLGHGPIR 162 MVLAILAFLRFTAIKPSLGLINRWGSVGKKEAMEIIKKFKKDLAAM LRIINARKERKRRGADTSIGIIGLLLTTAMAAEITRRGSAYYMYLDR SDAGKAISFATNLGVNKCHVQIMDLGHMCDATMSYECPMLDEGV EPDDVDCWCNTTSTWVVYGTCHHKKGEARRSRRAVTLPSHSTRKL QTRSQTWLESREYTKHLIKVENWIFRNPGFALAAVAIAWLLGSSTS QKVIYLVMILLIAPAYSIRCIGVSNRDFVEGMSGGTWVDVVLEHGG CVTVMAQDKPTVDIELVTTTVSNMAEVRSYCYEASISDMASDSRCP TQGEAYLDKQSDTQYVCKRTLVDRGWGNGCGLFGKGSLVTCAKF TCSKKMTGKSIQPENLEYRIMLSVHGSQHSGMIVNDENRAKVEVTP NSPRAEATLGGFGSLGLDCEPRTGLDFSDLYYLTMNNKHWLVHKE WFHDIPLPWHAGADTGTPHWNNKEALVEFKDAHAKRQTVVVLGS QEGAVHTALAGALEAEMDGAKGRLFSGHLKCRLKMDKLRLKGVS YSLCTAAFTFTKVPAETLHGTVTVEVQYAGTDGPCKVPAQMAVD MQTLTPVGRLITANPVITESTENSKMMLELDPPFGDSYIVIGVGDKK ITHHWHRSGSTIGKAFEATVRGAKRMAVLGDTAWDFGSVGGVFNS LGKGIHQIFGAAFKSLFGGMSWFSQILIGTLLVWLGLNTKNGSISLT CLALGGVMIFLSTAVSA ARB13565|AHF49783 MKNPKKKSGGFRIVNMLKRGVARVNPLGGLKRLPAGLLLGHGPIR 163 MVLAILAFLRFTAIKPSLGLINRWGSVGKKEAMEIIKKFKKDLAAM LRIINARKERKRRGADTSIGIIGLLLTTAMAAEITRRGSAYYMYLDR SDAGKAISFATNLGVNKCHVQIMDLGHMCDATMSYECPMLDEGV EPDDVDCWCNTTSTWVVYGTCHHKKGEARRSRRAVTLPSHSTRKL QTRSQTWLESREYTKHLIKVENWIFRNPGFALAAVAIAWLLGSSTS QKVIYLVMILLIAPAYSIRCIGVSNRDFVEGMSGGTWVDVVLEHGG CVTVMAQDKPTVDIELVTTTVSNMAEVRSYCYEASISDMASDSRCP TQGEAYLDKQSDTQYVCKRTLVDRGWGNGCGLFGKGSLVTCAKF TCSKKMTGKSIQPENLEYRIMLSVHGSQHSGMIVNDIGHETDENRA KVEVTPNSPRAEATLGGFGSLGLDCEPRTGLDFSDLYYLTMNNKH WLVHKEWFHDIPLPWHAGADTGTPHWNNKEALVEFKDAHAKRQT VVVLGSQEGAVHTALAGALEAEMDGAKGRLFSGHLKCRLKMDKL RLKGVSYSLCTAAFTFTKVPAETLHGTVTVEVQYAGTDGPCKVPA QMAVDMQTLTPVGRLITANPVITESTENSKMMLELDPPFGDSYIVIG VGDKKITHHWHRSGSTIGKAFEATVRGAKRMAVLGDTAWDFGSV GGVFNSLGKGVHQIFGAAFKSLFGGMSWFSQILIGTLLVWLGLNTK NGSISLTCLALGGVMIFLSTAVSA ArB1362|AHL43500 MKNPKKKSGGFRIVNMLKRGVARVNPLGGLKRLPAGLLLGHGPIR 164 MVLAILAFLRFTAIKPSLGLINRWGSVGKKEAMEIIKKFKKDLAAM LRIINARKERKRRGADTSIGIIGLLLTTAMAAEITRRGSAYYMYLDR SDAGKAISFATTLGVNKCHVQIMDLGHMCDATMSYECPMLDEGVE PDDVDCWCNTTSTWVVYGTCHHKKGEARRSRAVTLPSHSTRKL QTRSQTWLESREYTKHLIKVENWIFRNPGFALAAVAIAWLLGSSTS QKVIYLIMILLIAPAYSIRCIGVSNRDFVEGMSGGTWVDVVLEHGGC VTVMAQDKPTVDIELVTTTVSNMAEVRSYCYEASISDMASDSRCPT QGEAYLDKQSDTQYVCKRTLVDRGWGNGCGLFGKGSLVTCAKFT CSKKMTGKSIQPENLEYRIMLSVHGSQHSGMIVNDXXXXXXXNRA EVEVTPNSPRAEATLGGFGSLGLDCEPRTGLDFSDLYYLTMNNKH WLVHKEWFHDIPLPWHAGADTGTPHWNNKEALVEFKDAHAKRQT VVVLGSQEGAVHTALAGALEAEMDGAKGRLFSGHLKCRLKMDKL RLKGVSYSLCTAAFTFTKVPAETLHGTVTVEVQYAGTDGPCKVPA QMAVDMQTLTPVGRLITANPVITESTENSKMMLELDPPFGDSYIVIG VGDKKITHHWHRSGSTIGKAFEATVRGAKRMAVLGDTAWDFGSV GGVFNSLGKGIHQIFGAAFKSLFGGMSWFSQILIGTLLVWLGLNTK NGSISLTCLALGGVMIFLSTAVSA ArD7117|AHL43501 MKNPKKRSGGFRIVNMLKRGVARVNPLGGLKRLPAGLLLGHGPIR 165 MVLAILAFLRFTAIKPSLGLINRWGSVGKKEAMEIIKKFKKDLAAM LRIINARKERKRRGADTSIGIVGLLLTTAMAAEITRRGSAYYMYLDR SDAGKAISFATTLGVNKCHVQIMDLGHMCDATMSYECPMLDEGVE PDDVDCWCNTTSTWVVYGTCQHKKGEARRSRRAVTLPSHSTRKL QTRSQTWLESREYTKHLIKVENWIFRNPGFALVAVAIAWLLGSSTS QKVIYLVMILLIAPAYSIRCIGVSNRDFVEGMSGGTWVDVVLEHGG CVTVMAQDKPTVDIELVTTTVSNMAEVRSYCYEASISDMASDSRCP TQGEAYLDKQSDTQYVCKRTLVDRGWGNGCGLFGKGSLVTCAKF TCSKKMTGKSIQPENLEYRIMLSVHGSQHSGMIVNDIGHETDENRA KVEVTPNSPRAEATLGGFGSLGLDCEPRTGLDFSDLYYLTMNNKH WLVHKEWFHDIPLPWHAGADTGTPHWNNKEALVEFKDAHAKRQT VVVLGSQEGAVHTALAGALEAEMDGAKGRLFSGHLKCRLKMDKL RLKGVSYSLCTAVCTAAKVPAETLHGTVTVEVQYAGTDGPCKVPA QMAVDMQTLTPVGRLITANPVITESTENSKMMLELDPPFGDSYIVIG VGDKKITHHWHRSGSTIGKAFEATVRGAKRMAVLGDTAWDFGSV GGVFNSLGKGIHQIFGAAFKSLFGGMSWFSQILIGTLLVWLGLNTK NGSISLTCLALGGVMIFLSTAVSA ArD157995|AHL43503 MKNPKKKSGRFRIVNMLKRGVARVNPLGGLKRLPAGLLLGHGPIR 166 MVLAILAFLRFTAIKPSLGLINRWGSVGKKEAMEIIKKFKKDLAAM LRIINARKERKRRGADTSIGIIGLLLTTAMAAEITRRGSAYYMYLDR SDAGKAISFATTLGVNKCHVQIMDLGHMCDATMSYECPMLDEGVE PDDVDCWCNTTSTWVVYGTCHHKKGETRRSRRSVSLRYHYTRKL QTRSQTWLESREYKKHLIMVENWIFRNPGFAIVSVAITWLMGSLTS QKVIYLVMIVLIVPAYSISCIGVSNRDLVEGMSGGTWVDVVLEHGG CVTEMAQDKPTVDIELVTMTVSNMAEVRSYCYEASLSDMASASRC PTQGEPSLDKQSDTQSVCKRTLGDRGWGNGCGIFGKGSLVTCSKFT CCKKMPGKSIQPENLEYRIMLPVHGSQHSGMIVNDIGHETDENRAK VEVTPNSPRAEATLGGFGSLGLDCEPRTGLDFSDLYYLTMNNKHW LVHKEWFHDIPLPWHAGADTGTPHWNNKEALVEFKDAHAKRQTV VVLGSQEGAVHTALAGALEAEMDGAKGRLFSGHLKCRLKMDKLR LKGVSYSLCTAAFTFTKVPAETLHGTVTVEVQSAGTDGPCKVPAQ MAVDMQTLTPVGRLITANPVITESTENSKMMLELDPPFGDSYIVIGV GDKKITHHWHRSGSTIGKAFEATVRGAKRMAVLGDTAWDFGSVG GVFNSLGKGIHQIFGAAFKSLFGGMSWFSQILIGTLLVWLGLNTKN GSISLTCLALGGVMIFLSTAVSA ArD128000|AHL43502 MKNPKRKSGGFRIVNMLKRGVARVNPLGGLKRLPAGLLLGHGPIR 167 MVLAILAFLRFTAIKPSLGLINRWGSVGKKEAMEIIKKFKKDLAAM LRIINARKERKRRGADTSIGIIGLLLTTAMAAEITRRGSAYYMYLDR SDAGKAISFATTLGVNKCHVQIMDLGHMCDATMSYECPMLDEGVE PDDVDCWCNTTSTWVVYGTCHHKKGEARRSRRAVTLPSHSTRKL QTRSQTWLESREYTKHLIKVENWIFRNPGFALAAVAIAWLLGSSTS QKVIYLVMILLIAPAYSIRCIGVSNRDFVEGMSGGTWVDVVLEHGG CVTVMAQDKPTVDIELVTTTVSNMAEVRSYCYEASISDMASDSRCP TQGEAYLDKQSDTQYVCKRTLVDRGWGNGCGLFGKGSLVTCAKF TCSKKMTGKSIQPENLEYRIMLSVHGSQHSGMXXXXXGHETDENR AKVEVTPNSPRAEATLGGFGSLGLDCEPRTGLDFSDLYYLTMNNK HRLVRKEWFHDIPLPWHAGADTGTPHWNNKEALVEFKDAHAKRQ TVVVLGSQEGAVHTALAGALEAEMDGAKGRLFSGHLKCRLKMDK LRLKGVSYSLCTAAFTFTKVPAETLHGTVTVEVQYAGTDGPCKVP AQMAVDMQTLTPVGRLITANPVITESTENSKMMLELDPPFGDSYIVI GVGDKKITHHWLKKGSSIGKAFEATVRGAKRMAVLGDTAWDFGS VGGVFNSLGKGVHQIFGAAFKSLFGGMSWFSQILIGTLLVWLGLNT KNGSISLTCLALGGVMIFLSTAVSA ArD158084|AHL43504 MKNPKKKSGGFRIVNMLKRGVARVNPLGGLKRLPAGLLLGHGPIR 168 MVLAILAFLRFTAIKPSLGLINRWGSVGKKEAMEIIKKFKKDLAAM LRIINARKERKRRGADTSIGIIGLLLTTAMAAEITRRGSAYYMYLDR SDAGKAISFATTLGVNKCHVQIMDLGHMCDATMSYECPMLDEGVE PDDVDCWCNTTSTWVVYGTCHHKKGEARRSRRAVTLPSHSTRKL QTRSQTWLESREYTKHLIKVENWIFRNPGFALVAVAIAWLLGSSTS QKVIYLVMILLIAPAYSIRCIGVSNRDFVEGMSGGTWVDVVLEHGG CVTVMAQDKPTVDIELVTTTVSNMAEVRSYCYEASISDMASDSRCP TQGEAYLDKQSDTQYVCKRTLVDRGWGNGCGLFGKGSLVTCAKF TCSKKMTGKSIQPENLEYRIMLSVHGSQHSGMIVNDIGHETDENRA KVEVTPNSPRAEATLGGFGSLGLDCEPRTGLDFSDLYYLTMNNKH WLVHKEWFHDIPLPWHAGADTGTPHWNNKEALVEFKDAHAKRQT VVVLGSQEGAVHTALAGALEAEMDGAKGRLFSGHLKCRLKMDKL RLKGVSYSLCTAAFTFTKVPAETLHGTVTVEVQYAGTDGPCKVPA QMAVDMQTLTPVGRLITANPVITESTENSKMMLELDPPFGDSYIVIG VGDKKITHHWHRSGSTIGKAFEATVRGAKRMAVLGDTAWDFGSV GGVFNSLGKGIHQIFGAAFKSLFGGMSWFSQILIGTLLVWLGLNTK NGSISLTCLALGGVMIFLSTAVSA H/PF/2013|AHZ13508 MKNPKKKSGGFRIVNMLKRGVARVSPFGGLKRLPAGLLLGHGPIR 169 MVLAILAFLRFTAIKPSLGLINRWGSVGKKEAMEIIKKFKKDLAAM LRIINARKEKKRRGADTSVGIVGLLLTTAMAAEVTRRGSAYYMYL DRNDAGEAISFPTTLGMNKCYIQIMDLGHMCDATMSYECPMLDEG VEPDDVDCWCNTTSTWVVYGTCHHKKGEARRSRRAVTLPSHSTR KLQTRSQTWLESREYTKHLIRVENWIFRNPGFALAAAAIAWLLGSS TSQKVIYLVMILLIAPAYSIRCIGVSNRDFVEGMSGGTWVDVVLEH GGCVTVMAQDKPTVDIELVTTTVSNMAEVRSYCYEASISDMASDS RCPTQGEAYLDKQSDTQYVCKRTLVDRGWGNGCGLFGKGSLVTC AKFACSKKMTGKSIQPENLEYRIMLSVHGSQHSGMIVNDTGHETDE NRAKVEITPNSPRAEATLGGFGSLGLDCEPRTGLDFSDLYYLTMNN KHWLVHKEWFHDIPLPWHAGADTGTPHWNNKEALVEFKDAHAK RQTVVVLGSQEGAVHTALAGALEAEMDGAKGRLSSGHLKCRLKM DKLRLKGVSYSLCTAAFTFTKIPAETLHGTVTVEVQYAGTDGPCKV PAQMAVDMQTLTPVGRLITANPVITESTENSKMMLELDPPFGDSYI VIGVGEKKITHHWHRSGSTIGKAFEATVRGAKRMAVLGDTAWDFG SVGGALNSLGKGIHQIFGAAFKSLFGGMSWFSQILIGTLLMWLGLN TKNGSISLMCLALGGVLIFLSTAVSA MR766_NIID|BAP47441 MKNPKKKSGGFRIVNMLKRGVARVNPLGGLKRLPAGLLLGHGPIR 170 MVLAILAFLRFTAIKPSLGLINRWGSVGKKEAMEIIKKFKKDLAAM LRIINARKERKRRGADTSIGIIGLLLTTAMAAEITRRGSAYYMYLDR SDAGKAISFATTLGVNKCHVQIMDLGHMCDATMSYECPMLDEGVE PDDVDCWCNTTSTWVVYGTCHHKKGEARRSRRAVTLPSHSTRKL QTRSQTWLESREYTKHLIKVENWIFRNPGFALVAVAIAWLLGSSTS QKVIYLVMILLIAPAYSIRCIGVSNRDFVEGMSGGTWVDVVLEHGG CVTVMAQDKPTVDIELVTTTVSNMAEVRSYCYEASISDMASDSRCP TQGEAYLDKQSDTQYVCKRTLVDRGWGNGCGLFGKGSLVTCAKF TCSKKMTGKSIQPENLEYRIMLSVHGSQHSGMTVNDIGYETDENRA KVEVTPNSPRAEATLGGFGSLGLDCEPRTGLDFSDLYYLTMNNKH WLVHKEWFHDIPLPWHAGADTGTPHWNNKEALVEFKDAHAKRQT VVVLGSQEGAVHTALAGALEAEMDGAKGKLFSGHLKCRLKMDKL RLKGVSYSLCTAAFTFTKVPAETLHGTVTVEVQYAGTDGPCKIPVQ MAVDMQTLTPVGRLITANPVITESTENSKMMLELDPPFGDSYIVIGV GDKKITHHWHRSGSTIGKAFEATVRGAKRMAVLGDTAWDFGSVG GVFNSLGKGIHQIFGAAFKSLFGGMSWFSQILIGTLLVWLGLNTKN GSISLTCLALGGVMIFLSTAVSA prME VEVTKKGDTYYMFADKKDAGKVVTFETESGPNRCSIQAMDIGHM 171 ABI54480_South Africa CPATMSYECPVLEPQYEPEDVDCWCNSTAAWIVYGTCTHKTTGET RRSRRSITLPSHASQKLETRSSTWLESREYSKYLIKVENWILRNPGY ALVAAVIGWTLGSSRSQKIIFVTLLMLVAPAYSIRCIGIGNRDFIEGM SGGTWVDIVLEHGGCVTVMSNDKPTLDFELVTTTASNMAEVRSYC YEANISEMASDSRCPTQGEAYLDKMADSQFVCKRGYVDRGWGNG CGLFGKGSIVTCAKFTCVKKLTGKSIQPENLEYRVLVSVHASQHGG MINNDTNHQHDKENRARIDITASAPRVEVELGSFGSFSMECEPRSGL NFGDLYYLTMNNKHWLVNRDWFHDLSLPWHTGATSNNHHWNNK EALVEFREAHAKKQTAVVLGSQEGAVHAALAGALEAESDGHKATI YSGHLKCRLKLDKLRLKGMSYALCTGAFTFARTPSETIHGTATVEL QYAGEDGPCKVPIVITSDTNSMASTGRLITANPVVTESGANSKMMV EIDPPFGDSYIIVGTGTTKITHHWHRAGSSIGRAFEATMRGAKRMAV LGDTAWDFGSVGGMFNSVGKFVHQVFGSAFKALFGGMSWFTQLLI GFLLIWMGLNARGGTVAMSFMGIGAMLIFLATSVSG prME AEITRRGSAYYMYLDRSDAGKAISFATTLGVNKCHVQIMDLGHMC 172 AAV34151_Uganda_NHP DATMSYECPMLDEGVEPDDVDCWCNTTSTWVVYGTCHHKKGEA RRSRRAVTLPSHSTRKLQTRSQTWLESREYTKHLIKVENWIFRNPGF ALVAVAIAWLLGSSTSQKVIYLVMILLIAPAYSIRCIGVSNRDFVEG MSGGTWVDVVLEHGGCVTVMAQDKPTVDIELVTTTVSNMAEVRS YCYEASISDMASDSRCPTQGEAYLDKQSDTQYVCKRTLVDRGWGN GCGLFGKGSLVTCAKFTCSKKMTGKSIQPENLEYRIMLSVHGSQHS GMIGYETDEDRAKVEVTPNSPRAEATLGGFGSLGLDCEPRTGLDFS DLYYLTMNNKHWLVHKEWFHDIPLPWHAGADTGTPHWNNKEAL VEFKDAHAKRQTVVVLGSQEGAVHTALAGALEAEMDGAKGRLFS GHLKCRLKMDKLRLKGVSYSLCTAAFTFTKVPAETLHGTVTVEVQ YAGTDGPCKIPVQMAVDMQTLTPVGRLITANPVITESTENSKMMLE LDPPFGDSYIVIGVGDKKITHHWHRSGSTIGKAFEATVRGAKRMAV LGDTAWDFGSVGGVFNSLGKGIHQIFGAAFKSLFGGMSWFSQILIG TLLVWLGLNTKNGSISLTCLALGGVMIFLSTAVSA prME AEVTRRGSAYYMYLDRNDAGEAISFPTTLGMNKCYIQIMDLGHMC 173 AHZ13508_FrenchPoly_2013 DATMSYECPMLDEGVEPDDVDCWCNTTSTWVVYGTCHHKKGEA RRSRRAVTLPSHSTRKLQTRSQTWLESREYTKHLIRVENWIFRNPGF ALAAAAIAWLLGSSTSQKVIYLVMILLIAPAYSIRCIGVSNRDFVEG MSGGTWVDVVLEHGGCVTVMAQDKPTVDIELVTTTVSNMAEVRS YCYEASISDMASDSRCPTQGEAYLDKQSDTQYVCKRTLVDRGWGN GCGLFGKGSLVTCAKFACSKKMTGKSIQPENLEYRIMLSVHGSQHS GMIVNDTGHETDENRAKVEITPNSPRAEATLGGFGSLGLDCEPRTG LDFSDLYYLTMNNKHWLVHKEWFHDIPLPWHAGADTGTPHWNN KEALVEFKDAHAKRQTVVVLGSQEGAVHTALAGALEAEMDGAKG RLSSGHLKCRLKMDKLRLKGVSYSLCTAAFTFTKIPAETLHGTVTV EVQYAGTDGPCKVPAQMAVDMQTLTPVGRLITANPVITESTENSK MMLELDPPFGDSYIVIGVGEKKITHHWHRSGSTIGKAFEATVRGAK RMAVLGDTAWDFGSVGGALNSLGKGIHQIFGAAFKSLFGGMSWFS QILIGTLLMWLGLNTKNGSISLMCLALGGVLIFLSTAVSA prME AEITRRGSAYYMYLDRSDAGKAISFATTLGVNKCHVQIMDLGHMC 174 gAHL43504 DATMSYECPMLDEGVEPDDVDCWCNTTSTWVVYGTCHHKKGEA RRSRRAVTLPSHSTRKLQTRSQTWLESREYTKHLIKVENWIFRNPGF ALVAVAIAWLLGSSTSQKVIYLVMILLIAPAYSIRCIGVSNRDFVEG MSGGTWVDVVLEHGGCVTVMAQDKPTVDIELVTTTVSNMAEVRS YCYEASISDMASDSRCPTQGEAYLDKQSDTQYVCKRTLVDRGWGN GCGLFGKGSLVTCAKFTCSKKMTGKSIQPENLEYRIMLSVHGSQHS GMIVNDIGHETDENRAKVEVTPNSPRAEATLGGFGSLGLDCEPRTG LDFSDLYYLTMNNKHWLVHKEWFHDIPLPWHAGADTGTPHWNN KEALVEFKDAHAKRQTVVVLGSQEGAVHTALAGALEAEMDGAKG RLFSGHLKCRLKMDKLRLKGVSYSLCTAAFTFTKVPAETLHGTVTV EVQYAGTDGPCKVPAQMAVDMQTLTPVGRLITANPVITESTENSK MMLELDPPFGDSYIVIGVGDKKITHHWHRSGSTIGKAFEATVRGAK RMAVLGDTAWDFGSVGGVFNSLGKGIHQIFGAAFKSLFGGMSWFS QILIGTLLVWLGLNTKNGSISLTCLALGGVMIFLSTAVSA prME AEITRRGSAYYMYLDRSDAGKAISFATTLGVNKCHVQIMDLGHMC 175 AHL43503 DATMSYECPMLDEGVEPDDVDCWCNTTSTWVVYGTCHHKKGETR RSRRSVSLRYHYTRKLQTRSQTWLESREYKKHLIMVENWIFRNPGF AIVSVAITWLMGSLTSQKVIYLVMIVLIVPAYSISCIGVSNRDLVEG MSGGTWVDVVLEHGGCVTEMAQDKPTVDIELVTMTVSNMAEVRS YCYEASLSDMASASRCPTQGEPSLDKQSDTQSVCKRTLGDRGWGN GCGIFGKGSLVTCSKFTCCKKMPGKSIQPENLEYRIMLPVHGSQHSG MIVNDIGHETDENRAKVEVTPNSPRAEATLGGFGSLGLDCEPRTGL DFSDLYYLTMNNKHWLVHKEWFHDIPLPWHAGADTGTPHWNNK EALVEFKDAHAKRQTVVVLGSQEGAVHTALAGALEAEMDGAKGR LFSGHLKCRLKMDKLRLKGVSYSLCTAAFTFTKVPAETLHGTVTVE VQSAGTDGPCKVPAQMAVDMQTLTPVGRLITANPVITESTENSKM MLELDPPFGDSYIVIGVGDKKITHHWHRSGSTIGKAFEATVRGAKR MAVLGDTAWDFGSVGGVFNSLGKGIHQIFGAAFKSLFGGMSWFSQ ILIGTLLVWLGLNTKNGSISLTCLALGGVMIFLSTAVSA prME AAEITRRGSAYYMYLDRSDAGKAISFATTLGVNKCHVQIMDLGHM 176 AHL43502 CDATMSYECPMLDEGVEPDDVDCWCNTTSTWVVYGTCHHKKGE ARRSRRAVTLPSHSTRKLQTRSQTWLESREYTKHLIKVENWIFRNP GFALAAVAIAWLLGSSTSQKVIYLVMILLIAPAYSIRCIGVSNRDFVE GMSGGTWVDVVLEHGGCVTVMAQDKPTVDIELVTTTVSNMAEVR SYCYEASISDMASDSRCPTQGEAYLDKQSDTQYVCKRTLVDRGWG NGCGLFGKGSLVTCAKFTCSKKMTGKSIQPENLEYRIMLSVHGSQH SGMXXXXXGHETDENRAKVEVTPNSPRAEATLGGFGSLGLDCEPR TGLDFSDLYYLTMNNKHRLVRKEWFHDIPLPWHAGADTGTPHWN NKEALVEFKDAHAKRQTVVVLGSQEGAVHTALAGALEAEMDGAK GRLFSGHLKCRLKMDKLRLKGVSYSLCTAAFTFTKVPAETLHGTVT VEVQYAGTDGPCKVPAQMAVDMQTLTPVGRLITANPVITESTENS KMMLELDPPFGDSYIVIGVGDKKITHHWLKKGSSIGKAFEATVRGA KRMAVLGDTAWDFGSVGGVFNSLGKGVHQIFGAAFKSLFGGMSW FSQILIGTLLVWLGLNTKNGSISLTCLALGGVMIFLSTAVSA prME AEITRRGSAYYMYLDRSDAGKAISFATTLGVNKCHVQIMDLGHMC 177 AHL43501 DATMSYECPMLDEGVEPDDVDCWCNTTSTWVVYGTCQHKKGEA RRSRRAVTLPSHSTRKLQTRSQTWLESREYTKHLIKVENWIFRNPGF ALVAVAIAWLLGSSTSQKVIYLVMILLIAPAYSIRCIGVSNRDFVEG MSGGTWVDVVLEHGGCVTVMAQDKPTVDIELVTTTVSNMAEVRS YCYEASISDMASDSRCPTQGEAYLDKQSDTQYVCKRTLVDRGWGN GCGLFGKGSLVTCAKFTCSKKMTGKSIQPENLEYRIMLSVHGSQHS GMIVNDIGHETDENRAKVEVTPNSPRAEATLGGFGSLGLDCEPRTG LDFSDLYYLTMNNKHWLVHKEWFHDIPLPWHAGADTGTPHWNN KEALVEFKDAHAKRQTVVVLGSQEGAVHTALAGALEAEMDGAKG RLFSGHLKCRLKMDKLRLKGVSYSLCTAVCTAAKVPAETLHGTVT VEVQYAGTDGPCKVPAQMAVDMQTLTPVGRLITANPVITESTENS KMMLELDPPFGDSYIVIGVGDKKITHHWHRSGSTIGKAFEATVRGA KRMAVLGDTAWDFGSVGGVFNSLGKGIHQIFGAAFKSLFGGMSWF SQILIGTLLVWLGLNTKNGSISLTCLALGGVMIFLSTAVSA prME AEITRRGSAYYMYLDRSDAGKAISFATTLGVNKCHVQIMDLGHMC 178 AHL43500 DATMSYECPMLDEGVEPDDVDCWCNTTSTWVVYGTCHHKKGEA RRSRRAVTLPSHSTRKLQTRSQTWLESREYTKHLIKVENWIFRNPGF ALAAVAIAWLLGSSTSQKVIYLIMILLIAPAYSIRCIGVSNRDFVEGM SGGTWVDVVLEHGGCVTVMAQDKPTVDIELVTTTVSNMAEVRSY CYEASISDMASDSRCPTQGEAYLDKQSDTQYVCKRTLVDRGWGNG CGLFGKGSLVTCAKFTCSKKMTGKSIQPENLEYRIMLSVHGSQHSG MIVNDXXXXXXXNRAEVEVTPNSPRAEATLGGFGSLGLDCEPRTG LDFSDLYYLTMNNKHWLVHKEWFHDIPLPWHAGADTGTPHWNN KEALVEFKDAHAKRQTVVVLGSQEGAVHTALAGALEAEMDGAKG RLFSGHLKCRLKMDKLRLKGVSYSLCTAAFTFTKVPAETLHGTVTV EVQYAGTDGPCKVPAQMAVDMQTLTPVGRLITANPVITESTENSK MMLELDPPFGDSYIVIGVGDKKITHHWHRSGSTIGKAFEATVRGAK RMAVLGDTAWDFGSVGGVFNSLGKGIHQIFGAAFKSLFGGMSWFS QILIGTLLVWLGLNTKNGSISLTCLALGGVMIFLSTAVSA prME AEITRRGSAYYMYLDRSDAGKAISFATNLGVNKCHVQIMDLGHMC 179 AHF49785 DATMSYECPMLDEGVEPDDVDCWCNTTSTWVVYGTCHHKKGEA RRSRRAVTLPSHSTRKLQTRSQTWLESREYTKHLIKVENWIFRNPGF ALAAVAIAWLLGSSTSQKVIYLVMILLIAPAYSIRCIGVSNRDFVEG MSGGTWVDVVLEHGGCVTVMAQDKPTVDIELVTTTVSNMAEVRS YCYEASISDMASDSRCPTQGEAYLDKQSDTQYVCKRTLVDRGWGN GCGLFGKGSLVTCAKFTCSKKMTGKSIQPENLEYRIMLSVHGSQHS GMIVNDIGHETDENRAKVEVTPNSPRAEATLGGFGSLGLDCEPRTG LDFSDLYYLTMNNKHWLVHKEWFHDIPLPWHAGADTGTPHWNN KEALVEFKDAHAKRQTVVVLGSQEGAVHTALAGALEAEMDGAKG RLFSGHLKCRLKMDKLRLKGVSYSLCTAAFTFTKVPAETLHGTVTV EVQYAGTDGPCKVPAQMAVDMQTLTPVGRLITANPVITESTENSK MMLELDPPFGDSYIVIGVGDKKITHHWHRSGSTIGKAFEATVRGAK RMAVLGDTAWDFGSVGGVFNSLGKGVHQIFGAAFKSLFGGMSWF SQILIGTLLVWLGLNTKNGSISLTCLALGGVMIFLSTAVSA prME AEITRRGSAYYMYLDRSDAGKAISFATNLGVNKCHVQIMDLGHMC 180 AHF49784_1976 DATMSYECPMLDEGVEPDDVDCWCNTTSTWVVYGTCHHKKGEA RRSRRAVTLPSHSTRKLQTRSQTWLESREYTKHLIKVENWIFRNPGF ALAAVAIAWLLGSSTSQKVIYLVMILLIAPAYSIRCIGVSNRDFVEG MSGGTWVDVVLEHGGCVTVMAQDKPTVDIELVTTTVSNMAEVRS YCYEASISDMASDSRCPTQGEAYLDKQSDTQYVCKRTLVDRGWGN GCGLFGKGSLVTCAKFTCSKKMTGKSIQPENLEYRIMLSVHGSQHS GMIVNDENRAKVEVTPNSPRAEATLGGFGSLGLDCEPRTGLDFSDL YYLTMNNKHWLVHKEWFHDIPLPWHAGADTGTPHWNNKEALVE FKDAHAKRQTVVVLGSQEGAVHTALAGALEAEMDGAKGRLFSGH LKCRLKMDKLRLKGVSYSLCTAAFTFTKVPAETLHGTVTVEVQYA GTDGPCKVPAQMAVDMQTLTPVGRLITANPVITESTENSKMMLEL DPPFGDSYIVIGVGDKKITHHWHRSGSTIGKAFEATVRGAKRMAVL GDTAWDFGSVGGVFNSLGKGIHQIFGAAFKSLFGGMSWFSQILIGT LLVWLGLNTKNGSISLTCLALGGVMIFLSTAVSA prME AEITRRGSAYYMYLDRSDAGKAISFATNLGVNKCHVQIMDLGHMC 181 AHF49783 DATMSYECPMLDEGVEPDDVDCWCNTTSTWVVYGTCHHKKGEA RRSRRAVTLPSHSTRKLQTRSQTWLESREYTKHLIKVENWIFRNPGF ALAAVAIAWLLGSSTSQKVIYLVMILLIAPAYSIRCIGVSNRDFVEG MSGGTWVDVVLEHGGCVTVMAQDKPTVDIELVTTTVSNMAEVRS YCYEASISDMASDSRCPTQGEAYLDKQSDTQYVCKRTLVDRGWGN GCGLFGKGSLVTCAKFTCSKKMTGKSIQPENLEYRIMLSVHGSQHS GMIVNDIGHETDENRAKVEVTPNSPRAEATLGGFGSLGLDCEPRTG LDFSDLYYLTMNNKHWLVHKEWFHDIPLPWHAGADTGTPHWNN KEALVEFKDAHAKRQTVVVLGSQEGAVHTALAGALEAEMDGAKG RLFSGHLKCRLKMDKLRLKGVSYSLCTAAFTFTKVPAETLHGTVTV EVQYAGTDGPCKVPAQMAVDMQTLTPVGRLITANPVITESTENSK MMLELDPPFGDSYIVIGVGDKKITHHWHRSGSTIGKAFEATVRGAK RMAVLGDTAWDFGSVGGVFNSLGKGVHQIFGAAFKSLFGGMSWF SQILIGTLLVWLGLNTKNGSISLTCLALGGVMIFLSTAVSA prME VEVTRRGSAYYMYLDRSDAGEAISFPTTLGMNKCYIQIMDLGHMC 182 ACD75819_Micronesia DATMSYECPMLDEGVEPDDVDCWCNTTSTWVVYGTCHHKKGEA RRSRRAVTLPSHSTRKLQTRSQTWLESREYTKHLIRVENWIFRNPGF ALAAAAIAWLLGSSTSQKVIYLVMILLIAPAYSIRCIGVSNRDFVEG MSGGTWVDVVLEHGGCVTVMAQDKPAVDIELVTTTVSNMAEVRS YCYEASISDMASDSRCPTQGEAYLDKQSDTQYVCKRTLVDRGWGN GCGLFGKGSLVTCAKFACSKKMTGKSIQPENLEYRIMLSVHGSQHS GMIVNDTGHETDENRAKVEITPNSPRAEATLGGFGSLGLDCEPRTG LDFSDLYYLTMNNKHWLVHKEWFHDIPLPWHAGADTGTPHWNN KEALVEFKDAHAKRQTVVVLGSQEGAVHTALAGALEAEMDGAKG RLSSGHLKCRLKMDKLRLKGVSYSLCTAAFTFTKIPAETLHGTVTV EVQYAGTDGPCKVPAQMAVDMQTLTPVGRLITANPVITESTENSK MMLELDPPFGDSYIVIGVGEKKITHHWHRSGSTIGKAFEATVRGAK RMAVLGDTAWDFGSVGGALNSLGKGIHQIFGAAFKSLFGGMSWFS QILIGTLLVWLGLNTKNGSISLTCLALGGVLIFLSTAVSA prME AEITRRGSAYYMYLDRSDAGKAISFATTLGVNKCHVQIMDLGHMC 183 ABI54475 DATMSYECPMLDEGVEPDDVDCWCNTTSTWVVYGTCHHKKGEA RRSRRAVTLPSHSTRKLQTRSQTWLESREYTKHLIKVENWIFRNPGF TLVAVAIAWLLGSSTSQKVIYLVMILLIAPAYSIRCIGVSNRDFVEG MSGGTWVDVVLEHGGCVTVMAQDKPTVDIELVTTTVSNMAEVRS YCYEASISDMASDSRCPTQGEAYLDKQSDTQYVCKRTLVDRGWGN GCGLFGKGSLVTCAKFTCSKKMTGKSIQPENLEYRIMLSVHGSQHS GMIVNDENRAKVEVTPNSPRAEATLGGFGSLGLDCEPRTGLDFSDL YYLTMNNKHWLVHKEWFHDIPLPWHAGADTGTPHWNNKEALVE FKDAHAKRQTVVVLGSQEGAVHTALAGALEAEMDGAKGRLFSGH LKCRLKMDKLRLKGVSYSLCTAAFTFTKVPAETLHGTVTVEVQYA GTDGPCKVPAQMAVDMQTLTPVGRLITANPVITESTENSKMMLEL DPPFGDSYIVIGVGDKKITHHWHRSGSTIGKAFEATVRGAKRMAVL GDTAWDFGSVGGVFNSLGKGIHQIFGAAFKSLFGGMSWFSQILIGT LLVWLGLNTKNGSISLTCLALGGVMIFLSTAVSA prME AEITRRGSAYYMYLDRSDAGKAISFATTLGVNKCHVQIMDLGHMC 184 YP_002790881 DATMSYECPMLDEGVEPDDVDCWCNTTSTWVVYGTCHHKKGEA RRSRRAVTLPSHSTRKLQTRSQTWLESREYTKHLIKVENWIFRNPGF ALVAVAIAWLLGSSTSQKVIYLVMILLIAPAYSIRCIGVSNRDFVEG MSGGTWVDVVLEHGGCVTVMAQDKPTVDIELVTTTVSNMAEVRS YCYEASISDMASDSRCPTQGEAYLDKQSDTQYVCKRTLVDRGWGN GCGLFGKGSLVTCAKFTCSKKMTGKSIQPENLEYRIMLSVHGSQHS GMIGYETDEDRAKVEVTPNSPRAEATLGGFGSLGLDCEPRTGLDFS DLYYLTMNNKHWLVHKEWFHDIPLPWHAGADTGTPHWNNKEAL VEFKDAHAKRQTVVVLGSQEGAVHTALAGALEAEMDGAKGRLFS GHLKCRLKMDKLRLKGVSYSLCTAAFTFTKVPAETLHGTVTVEVQ YAGTDGPCKIPVQMAVDMQTLTPVGRLITANPVITESTENSKMMLE LDPPFGDSYIVIGVGDKKITHHWHRSGSTIGKAFEATVRGAKRMAV LGDTAWDFGSVGGVFNSLGKGIHQIFGAAFKSLFGGMSWFSQILIG TLLVWLGLNTKNGSISLTCLALGGVMIFLSTAVSA prME AEITRRGSAYYMYLDRSDAGKAISFATTLGVNKCHVQIMDLGHMC 185 BAP4744 DATMSYECPMLDEGVEPDDVDCWCNTTSTWVVYGTCHHKKGEA RRSRRAVTLPSHSTRKLQTRSQTWLESREYTKHLIKVENWIFRNPGF ALVAVAIAWLLGSSTSQKVIYLVMILLIAPAYSIRCIGVSNRDFVEG MSGGTWVDVVLEHGGCVTVMAQDKPTVDIELVTTTVSNMAEVRS YCYEASISDMASDSRCPTQGEAYLDKQSDTQYVCKRTLVDRGWGN GCGLFGKGSLVTCAKFTCSKKMTGKSIQPENLEYRIMLSVHGSQHS GMTVNDIGYETDENRAKVEVTPNSPRAEATLGGFGSLGLDCEPRTG LDFSDLYYLTMNNKHWLVHKEWFHDIPLPWHAGADTGTPHWNN KEALVEFKDAHAKRQTVVVLGSQEGAVHTALAGALEAEMDGAKG KLFSGHLKCRLKMDKLRLKGVSYSLCTAAFTFTKVPAETLHGTVT VEVQYAGTDGPCKIPVQMAVDMQTLTPVGRLITANPVITESTENSK MMLELDPPFGDSYIVIGVGDKKITHHWHRSGSTIGKAFEATVRGAK RMAVLGDTAWDFGSVGGVFNSLGKGIHQIFGAAFKSLFGGMSWFS QILIGTLLVWLGLNTKNGSISLTCLALGGVMIFLSTAVSA prME AEVTRRGSAYYMYLDRNDAGEAISFPTTLGMNKCYIQIMDLGHMC 186 KU365780_2015_Brazil_ DATMSYECPMLDEGVEPDDVDCWCNTTSTWVVYGTCHHKKGEA isolate_BeH815744 RRSRRAVTLPSHSTRKLQTRSQTWLESREYTKHLIRVENWIFRNPGF ALAAAAIAWLLGSSTSQKVIYLVMILLIAPAYSIRCIGVSNRDFVEG MSGGTWVDVVLEHGGCVTVMAQDKPTVDIELVTTTVSNMAEVRS YCYEASISDMASDSRCPTQGEAYLDKQSDTQYVCKRTLVDRGWGN GCGLFGKGSLVTCAKFACSKKMTGKSIQPENLEYRIMLSVHGSQHS GMIVNDTGHETDENRAKVEITPNSPRAEATLGGFGSLGLDCEPRTG LDFSDLYYLTMNNKHWLVHKEWFHDIPLPWHAGADTGTPHWNN KEALVEFKDAHAKRQTVVVLGSQEGAVHTALAGALEAEMDGAKG RLSSGHLKCRLKMDKLRLKGVSYSLCTAAFTFTKIPAETLHGTVTV EVQYAGTDGPCKVPAQMAVDMQTLTPVGRLITANPVITESTENSK MMLELDPPFGDSYIVIGVGEKKITHHWHRSGSTIGKAFEATVRGAK RMAVLGDTAWDFGSVGGALNSLGKGIHQIFGAAFKSLFGGMSWFS QILIGTLLMWLGLNTKNGSISLMCLALGGVLIFLSTAVSA prME AEVTRRGSAYYMYLDRNDAGEAISFPTTLGMNKCYIQIMDLGHMC 187 KU365779_2015_Brazil_ DATMSYECPMLDEGVEPDDVDCWCNTTSTWVVYGTCHHKKGEA isolate_BeH819966 RRSRRAVTLPSHSTRKLQTRSQTWLESREYTKHLIRVENWIFRNPGF ALAAAAIAWLLGSSTSQKVIYLVMILLIAPAYSIRCIGVSNRDFVEG MSGGTWVDVVLEHGGCVTVMAQDKPTVDIELVTTTVSNMAEVRS YCYEASISDMASDSRCPTQGEAYLDKQSDTQYVCKRTLVDRGWGN GCGLFGKGSLVTCAKFACSKKMTGKSIQPENLEYRIMLSVHGSQHS GMIVNDTGHETDENRAKVEITPNSPRAEATLGGFGSLGLDCEPRTG LDFSDLYYLTMNNKHWLVHKEWFHDIPLPWHAGADTGTPHWNN KEALVEFKDAHAKRQTVVVLGSQEGAVHTALAGALEAEMDGAKG RLSSGHLKCRLKMDKLRLKGVSYSLCTAAFTFTKIPAETLHGTVTV EVQYAGTDGPCKVPAQMAVDMQTLTPVGRLITANPVITESTENSK MMLELDPPFGDSYIVIGVGEKKITHHWHRSGSTIGKAFEATVRGAK RMAVLGDTAWDFGSVGGALNSLGKGIHQIFGAAFKSLFGGMSWFS QILIGTLLMWLGLNTKNGSISLMCLALGGVLIFLSTAVSA prME AEVTRRGSAYYMYLDRNDAGEAISFPTTLGMNKCYIQIMDLGHMC 188 KU365778_2015_Brazil_ DATMSYECPMLDEGVEPDDVDCWCNTTSTWVVYGTCHHKKGEA isolate_BeH819015 RRSRRAVTLPSHSTRKLQTRSQTWLESREYTKHLIRVENWIFRNPGF ALAAAAIAWLLGSSTSQKVIYLVMILLIAPAYSIRCIGVSNRDFVEG MSGGTWVDVVLEHGGCVTVMAQDKPTVDIELVTTTVSNMAEVRS YCYEASISDMASDSRCPTQGEAYLDKQSDTQYVCKRTLVDRGWGN GCGLFGKGSLVTCAKFACSKKMTGKSIQPENLEYRIMLSVHGSQHS GMIVNDTGHETDENRAKVEITPNSPRAEATLGGFGSLGLDCEPRTG LDFSDLYYLTMNNKHWLVHKEWFHDIPLPWHAGADTGTPHWNN KEALVEFKDAHAKRQTVVVLGSQEGAVHTALAGALEAEMDGAKG RLSSGHLKCRLKMDKLRLKGVSYSLCTAAFTFTKIPAETLHGTVTV EVQYAGTDGPCKVPAQMAVDMQTLTPVGRLITANPVITESTENSK MMLELDPPFGDSYIVIGVGEKKITHHWHRSGSTIGKAFEATVRGAK RMAVLGDTAWDFGSVGGALNSLGKGIHQIFGAAFKSLFGGMSWFS QILIGTLLMWLGLNTKNGSISLMCLALGGVLIFLSTAVSA prME AEVTRRGSAYYMYLDRNDAGEAISFPTTLGMNKCYIQIMDLGHMC 189 KU365777_2015_Brazil_ DATMSYECPMLDEGVEPDDVDCWCNTTSTWVVYGTCHHKKGEA isolate_BeH818995 RRSRRAVTLPSHSTRKLQTRSQTWLESREYTKHLIRVENWIFRNPGF ALAAAAIAWLLGSSTSQKVIYLVMILLIAPAYSIRCIGVSNRDFVEG MSGGTWVDVVLEHGGCVTVMAQDKPTVDIELVTTTVSNMAEVRS YCYEASISDMASDSRCPTQGEAYLDKQSDTQYVCKRTLVDRGWGN GCGLFGKGSLVTCAKFACSKKMTGKSIQPENLEYRIMLSVHGSQHS GMIVNDTGHETDENRAKVEITPNSPRAEATLGGFGSLGLDCEPRTG LDFSDLYYLTMNNKHWLVHKEWFHDIPLPWHAGADTGTPHWNN KEALVEFKDAHAKRQTVVVLGSQEGAVHTALAGALEAEMDGAKG RLSSGHLKCRLKMDKLRLKGVSYSLCTAAFTFTKIPAETLHGTVTV EVQYAGTDGPCKVPAQMAVDMQTLTPVGRLITANPVITESTENSK MMLELDPPFGDSYIVIGVGEKKITHHWHRSGSTIGKAFEATVRGAK RMAVLGDTAWDFGSVGGALNSLGKGIHQIFGAAFKSLFGGMSWFS QILIGTLLMWLGLNTKNGSISLMCLALGGVLIFLSTAVSA prME AEVTRRGSAYYMYLDRNDAGEAISFPTTLGMNKCYIQIMDLGHMC 190 KU321639_2015_Brazil_ DATMSYECPMLDEGVEPDDVDCWCNTTSTWVVYGTCHHKKGEA isolate_ZikaSPH2015 RRSRRAVTLPSHSTRKLQTRSQTWLESREYTKHLIRVENWIFRNPGF ALAAAAIAWLLGSSTSQKVIYLVMILLIAPAYSIRCIGVSNRDFVEG MSGGTWVDIVLEHGGCVTVMAQDKPTVDIELVTTTVSNMAEVRS YCYEASISDMASDSRCPTQGEAYLDKQSDTQYVCKRTLVDRGWGN GCGLFGKGSLVTCAKFACSKKMTGKSIQPENLEYRIMLSVHGSQHS GMIVNDTGHETDENRAKVEITPNSPRAEATLGGFGSLGLDCEPRTG LDFSDLYYLTMNNKHWLVHKEWFHDIPLPWHAGADTGTPHWNN KEALVEFKDAHAKRQTVVVLGSQEGAVHTALAGALEAEMDGAKG RLSSGHLKCRLKMDKLRLKGVSYSLCTAAFTFTKIPAETLHGTVTV EVQYAGTDGPCKVPAQMAVDMQTLTPVGRLITANPVITESTENSK MMLELDPPFGDSYIVIGVGEKKITHHWHRSGSTIGKAFEATVRGAK RMAVLGDTAWDFGSVGGALNSLGKGIHQIFGAAFKSLFGGMSWFS QILIGTLLMWLGLNTKNGSISLMCLALGGVLIFLSTAVSA prME AEVTRRGSAYYMYLDRNDAGEAISFPTTLGMNKCYIQIMDLGHTC 191 KU312312_2015_Suriname_ DATMSYECPMLDEGVEPDDVDCWCNTTSTWVVYGTCHHKKGEA isolate_Z1106033 RRSRRAVTLPSHSTRKLQTRSQTWLESREYTKHLIRVENWIFRNPGF ALAAAAIAWLLGSSTSQKVIYLVMILLIAPAYSIRCIGVSNRDFVEG MSGGTWVDVVLEHGGCVTVMAQDKPTVDIELVTTTVSNMAEVRS YCYEASISDMASDSRCPTQGEAYLDKQSDTQYVCKRTLVDRGWGN GCGLFGKGSLVTCAKFACSKKMTGKSIQPENLEYRIMLSVHGSQHS GMIVNDTGHETDENRAKVEITPNSPRAEATLGGFGSLGLDCEPRTG LDFSDLYYLTMNNKHWLVHKEWFHDIPLPWHAGADTGTPHWNN KEALVEFKDAHAKRQTVVVLGSQEGAVHTALAGALEAEMDGAKG RLSSGHLKCRLKMDKLRLKGVSYSLCTAAFTFTKIPAETLHGTVTV EVQYAGTDGPCKVPAQMAVDMQTLTPVGRLITANPVITESTENSK MMLELDPPFGDSYIVIGVGEKKITHHWHRSGSTIGKAFEATVRGAK RMAVLGDTAWDFGSVGGALNSLGKGIHQIFGAAFKSLFGGMSWFS QILIGTLLMWLGLNAKNGSISLMCLALGGVLIFLSTAVSA Premembrane/membrane AEVTRRGSAYYMYLDRNDAGEAISFPTTLGMNKCYIQIMDLGHMC 469 protein DATMSYECPMLDEGVEPDDVDCWCNTTSTWVVYGTCHHKKGEA KU321639_2015_Brazil_ RRSRRAVTLPSHSTRKLQTRSQTWLESREYTKHLIRVENWIFRNPGF isolate_ZikaSPH2015 ALAAAAIAWLLGSSTSQKVIYLVMILLIAPAYS Envelop protein IRCIGVSNRDFVEGMSGGTWVDIVLEHGGCVTVMAQDKPTVDIEL 192 KU321639_2015_Brazil_ VTTTVSNMAEVRSYCYEASISDMASDSRCPTQGEAYLDKQSDTQY isolate_ZikaSPH2015 VCKRTLVDRGWGNGCGLFGKGSLVTCAKFACSKKMTGKSIQPENL EYRIMLSVHGSQHSGMIVNDTGHETDENRAKVEITPNSPRAEATLG GFGSLGLDCEPRTGLDFSDLYYLTMNNKHWLVHKEWFHDIPLPWH AGADTGTPHWNNKEALVEFKDAHAKRQTVVVLGSQEGAVHTALA GALEAEMDGAKGRLSSGHLKCRLKMDKLRLKGVSYSLCTAAFTFT KIPAETLHGTVTVEVQYAGTDGPCKVPAQMAVDMQTLTPVGRLIT ANPVITESTENSKMMLELDPPFGDSYIVIGVGEKKITHHWHRSGSTI GKAFEATVRGAKRMAVLGDTAWDFGSVGGALNSLGKGIHQIFGA AFKSLFGGMSWFSQILIGTLLMWLGLNTKNGSISLMCLALGGVLIFL STAVSA Capsid protein MKNPKKKSGGFRIVNMLKRGVARVSPFGGLKRLPAGLLLGHGPIR 193 KU321639_2015_Brazil_ MVLAILAFLRFTAIKPSLGLINRWGSVGKKEAMEIIKKFKKDLAAM isolate_ZikaSPH2015 LRIINARKEKKRRGADTSVGIVGLLLTTAMAAEV Non-structural protein 1 VGCSVDFSKKETRCGTGVFVYNDVEAWRDRYKYHPDSPRRLAAA 194 KU321639_2015_Brazil_ VKQAWEDGICGISSVSRMENIMWRSVEGELNAILEENGVQLTVVV isolate_ZikaSPH2015 GSVKNPMWRGPQRLPVPVNELPHGWKAWGKSHFVRAAKTNNSFV VDGDTLKECPLKHRAWNSFLVEDHGFGVFHTSVWLKVREDYSLEC DPAVIGTAVKGKEAVHSDLGYWIESEKNDTWRLKRAHLIEMKTCE WPKSHTLWTDGIEESDLIIPKSLAGPLSHHNTREGYRTQMKGPWHS EELEIRFEECPGTKVHVEETCGTRGPSLRSTTASGRVIEEWCCRECT MPPLSFRAKDGCWYGMEIRPRKEPESNLVRSMVTAGSTDHMDHFS L Non-structural protein 2A GVLVILLMVQEGLKKRMTTKIIISTSMAVLVAMILGGFSMSDLAKL 195 KU321639_2015_Brazil_ AILMGATFAEMNTGGDVAHLALIAAFKVRPALLVSFIFRANWTPRE isolate_ZikaSPH2015 SMLLALASCLLQTAISALEGDLMVLINGFALAWLAIRAMVVPRTDN ITLAILAALTPLARGTLLVAWRAGLATCGGFMLLSLKGKGSVKKNL PFVMALGLTAVRLVDPINVVGLLLLTRSGKRSWP Non-structural protein 2B PSEVLTAVGLICALAGGFAKADIEMAGPMAAVGLLIVSYVVSGKSV 196 KU321639_2015_Brazil_ DMYIERAGDITWEKDAEVTGNSPRLDVALDESGDFSLVEDDGPPM isolate_ZikaSPH2015 REIILKVVLMTICGMNPIAIPFAAGAWYVYVKTGKRSGALWDVPAP KEVKKGE Non-structural protein 3 TTDGVYRVMTRRLLGSTQVGVGVMQEGVFHTMWHVTKGSALRS 197 KU321639_2015_Brazil_ GEGRLDPYWGDVKQDLVSYCGPWKLDAAWDGHSEVQLLAVPPG isolate_ZikaSPH2015 ERARNIQTLPGIFKTKDGDIGAVALDYPAGTSGSPILDKCGRVIGLY GNGVVIKNGSYVSAITQGRREEETPVECFEPSMLKKKQLTVLDLHP GAGKTRRVLPEIVREAIKTRLRTVILAPTRVVAAEMEEALRGLPVR YMTTAVNVTHSGTEIVDLMCHATFTSRLLQPIRVPNYNLYIMDEAH FTDPSSIAARGYISTRVEMGEAAAIFMTATPPGTRDAFPDSNSPIMDT EVEVPERAWSSGFDWVTDYSGKTVWFVPSVRNGNEIAACLTKAGK RVIQLSRKTFETEFQKTKHQEWDFVVTTDISEMGANFKADRVIDSR RCLKPVILDGERVILAGPMPVTHASAAQRRGRIGRNPNKPGDEYLY GGGCAETDEDHAHWLEARMLLDNIYLQDGLIASLYRPEADKVAA IEGEFKLRTEQRKTFVELMKRGDLPVWLAYQVASAGITYTDRRWC FDGTTNNTIMEDSVPAEVWTRHGEKRVLKPRWMDARVCSDHAAL KSFKEFAAGKRGAA Non-structural protein 4A FGVMEALGTLPGHMTERFQEAIDNLAVLMRAETGSRPYKAAAAQL 198 KU321639_2015_Brazil_ PETLETIMLLGLLGTVSLGIFFVLMRNKGIGKMGFGMVTLGASAWL isolate_ZikaSPH2015 MWLSEIEPARIACVLIVVFLLLVVLIPEPEKQRSPQDNQMAIIIMVAV GLLGLITA Non-structural protein 4B NELGWLERTKSDLSHLMGRREEGATMGFSMDIDLRPASAWAIYAA 199 KU321639_2015_Brazil_ LTTFITPAVQHAVTTSYNNYSLMAMATQAGVLFGMGKGMPFYAW isolate_ZikaSPH2015 DFGVPLLMIGCYSQLTPLTLIVAIILLVAHYMYLIPGLQAAAARAAQ KRTAAGIMKNPVVDGIVVTDIDTMTIDPQVEKKMGQVLLMAVAVS SAILSRTAWGWGEAGALITAATSTLWEGSPNKYWNSSTATSLCNIF RGSYLAGASLIYTVTRNAGLVKRRGGGTGETLGEKWKARLNQMS ALEFYSYKKSGITEVCREEARRALKDGVATGGHAVSRGSAKLRWL VERGYLQPYGKVIDLGCGRGGWSYYAATIRKVQEVKGYTKGGPG HEEPVLVQSYGWNIVRLKSGVDVFHMAAEPCDTLLCDIGESSSSPE VEEARTLRVLSMVGDWLEKRPGAFCIKVLCPYTSTMMETLERLQR RYGGGLVRVPLSRNSTHEMYWVSGAKSNTIKSVSTTSQLLLGRMD GPRRPV Non-structural protein 5 KYEEDVNLGSGTRAVVSCAEAPNMKIIGNRIERIRSEHAETWFFDEN 200 KU321639_2015_Brazil_ HPYRTWAYHGSYEAPTQGSASSLINGVVRLLSKPWDVVTGVTGIA isolate_ZikaSPH2015 MTDTTPYGQQRVFKEKVDTRVPDPQEGTRQVMSMVSSWLWKELG KHKRPRVCTKEEFINKVRSNAALGAIFEEEKEWKTAVEAVNDPRF WALVDKEREHHLRGECQSCVYNMMGKREKKQGEFGKAKGSRAI WYMWLGARFLEFEALGFLNEDHWMGRENSGGGVEGLGLQRLGY VLEEMSRIPGGRMYADDTAGWDTRISRFDLENEALITNQMEKGHR ALALAIIKYTYQNKVVKVLRPAEKGKTVMDIISRQDQRGSGQVVTY ALNTFTNLVVQLIRNMEAEEVLEMQDLWLLRRSEKVTNWLQSNG WDRLKRMAVSGDDCVVKPIDDRFAHALRFLNDMGKVRKDTQEW KPSTGWDNWEEVPFCSHHFNKLHLKDGRSIVVPCRHQDELIGRAR VSPGAGWSIRETACLAKSYAQMWQLLYFHRRDLRLMANAICSSVP VDWVPTGRTTWSIHGKGEWMTTEDMLVVWNRVWIEENDHMEDK TPVTKWTDIPYLGKREDLWCGSLIGHRPRTTWAENIKNTVNMVRRI IGDEEKYMDYLSTQVRYLGEEGSTPGVL Signal peptide_prM-E METPAQLLFLLLLWLPDTTGAEVTRRGSAYYMYLDRNDAGEAISFP 201 TTLGMNKCYIQIMDLGHMCDATMSYECPMLDEGVEPDDVDCWCN TTSTWVVYGTCHHKKGEARRSRRAVTLPSHSTRKLQTRSQTWLES REYTKHLIRVENWIFRNPGFALAAAAIAWLLGSSTSQKVIYLVMILL IAPAYSIRCIGVSNRDFVEGMSGGTWVDIVLEHGGCVTVMAQDKPT VDIELVTTTVSNMAEVRSYCYEASISDMASDSRCPTQGEAYLDKQS DTQYVCKRTLVDRGWGNGCGLFGKGSLVTCAKFACSKKMTGKSI QPENLEYRIMLSVHGSQHSGMIVNDTGHETDENRAKVEITPNSPRA EATLGGFGSLGLDCEPRTGLDFSDLYYLTMNNKHWLVHKEWFHDI PLPWHAGADTGTPHWNNKEALVEFKDAHAKRQTVVVLGSQEGAV HTALAGALEAEMDGAKGRLSSGHLKCRLKMDKLRLKGVSYSLCT AAFTFTKIPAETLHGTVTVEVQYAGTDGPCKVPAQMAVDMQTLTP VGRLITANPVITESTENSKMMLELDPPFGDSYIVIGVGEKKITHHWH RSGSTIGKAFEATVRGAKRMAVLGDTAWDFGSVGGALNSLGKGIH QIFGAAFKSLFGGMSWFSQILIGTLLMWLGLNTKNGSISLMCLALG GVLIFLSTAVSA Signal peptide_E METPAQLLFLLLLWLPDTTGIRCIGVSNRDFVEGMSGGTWVDIVLE 202 HGGCVTVMAQDKPTVDIELVTTTVSNMAEVRSYCYEASISDMASD SRCPTQGEAYLDKQSDTQYVCKRTLVDRGWGNGCGLFGKGSLVT CAKFACSKKMTGKSIQPENLEYRIMLSVHGSQHSGMIVNDTGHETD ENRAKVEITPNSPRAEATLGGFGSLGLDCEPRTGLDFSDLYYLTMN NKHWLVHKEWFHDIPLPWHAGADTGTPHWNNKEALVEFKDAHA KRQTVVVLGSQEGAVHTALAGALEAEMDGAKGRLSSGHLKCRLK MDKLRLKGVSYSLCTAAFTFTKIPAETLHGTVTVEVQYAGTDGPCK VPAQMAVDMQTLTPVGRLITANPVITESTENSKMMLELDPPFGDSY IVIGVGEKKITHHWHRSGSTIGKAFEATVRGAKRMAVLGDTAWDF GSVGGALNSLGKGIHQIFGAAFKSLFGGMSWFSQILIGTLLMWLGL NTKNGSISLMCLALGGVLIFLSTAVSA IgE HC signal MDWTWILFLVAAATRVHSVEVTRRGSAYYMYLDRSDAGEAISFPT 203 peptide_prM-E #1 TLGMNKCYIQIMDLGHMCDATMSYECPMLDEGVEPDDVDCWCNT (Brazil_isolate_ TSTWVVYGTCHHKKGEARRSRRAVTLPSHSTRKLQTRSQTWLESR ZikaSPH2015) EYTKHLIRVENWIFRNPGFALAAAAIAWLLGSSTSQKVIYLVMILLI APAYSIRCIGVSNRDFVEGMSGGTWVDVVLEHGGCVTVMAQDKP AVDIELVTTTVSNMAEVRSYCYEASISDMASDSRCPTQGEAYLDKQ SDTQYVCKRTLVDRGWGNGCGLFGKGSLVTCAKFACSKKMTGKSI QPENLEYRIMLSVHGSQHSGMIVNDTGHETDENRAKVEITPNSPRA EATLGGFGSLGLDCEPRTGLDFSDLYYLTMNNKHWLVHKEWFHDI PLPWHAGADTGTPHWNNKEALVEFKDAHAKRQTVVVLGSQEGAV HTALAGALEAEMDGAKGRLSSGHLKCRLKMDKLRLKGVSYSLCT AAFTFTKIPAETLHGTVTVEVQYAGTDGPCKVPAQMAVDMQTLTP VGRLITANPVITESTENSKMMLELDPPFGDSYIVIGVGEKKITHHWH RSGSTIGKAFEATVRGAKRMAVLGDTAWDFGSVGGALNSLGKGIH QIFGAAFKSLFGGMSWFSQILIGTLLVWLGLNTKNGSISLTCLALGG VLIFLSTAVSA IgE HC signal MDWTWILFLVAAATRVHSVEVTRRGSAYYMYLDRSDAGEAISFPT 204 peptide_prM-E #1 TLGMNKCYIQIMDLGHMCDATMSYECPMLDEGVEPDDVDCWCNT (ACD75819_Micronesia) TSTWVVYGTCHHKKGEARRSRRAVTLPSHSTRKLQTRSQTWLESR EYTKHLIRVENWIFRNPGFALAAAAIAWLLGSSTSQKVIYLVMILLI APAYSIRCIGVSNRDFVEGMSGGTWVDVVLEHGGCVTVMAQDKP AVDIELVTTTVSNMAEVRSYCYEASISDMASDSRCPTQGEAYLDKQ SDTQYVCKRTLVDRGWGNGCGLFGKGSLVTCAKFACSKKMTGKSI QPENLEYRIMLSVHGSQHSGMIVNDTGHETDENRAKVEITPNSPRA EATLGGFGSLGLDCEPRTGLDFSDLYYLTMNNKHWLVHKEWFHDI PLPWHAGADTGTPHWNNKEALVEFKDAHAKRQTVVVLGSQEGAV HTALAGALEAEMDGAKGRLSSGHLKCRLKMDKLRLKGVSYSLCT AAFTFTKIPAETLHGTVTVEVQYAGTDGPCKVPAQMAVDMQTLTP VGRLITANPVITESTENSKMMLELDPPFGDSYIVIGVGEKKITHHWH RSGSTIGKAFEATVRGAKRMAVLGDTAWDFGSVGGALNSLGKGIH QIFGAAFKSLFGGMSWFSQILIGTLLVWLGLNTKNGSISLTCLALGG VLIFLSTAVSA IgE HC signal MDWTWILFLVAAATRVHSTRRGSAYYMYLDRSDAGEAISFPTTLG 205 peptide_prM-E #2 MNKCYIQIMDLGHMCDATMSYECPMLDEGVEPDDVDCWCNTTST (Brazil_isolate_ WVVYGTCHHKKGEARRSRRAVTLPSHSTRKLQTRSQTWLESREYT ZikaSPH2015) KHLIRVENWIFRNPGFALAAAAIAWLLGSSTSQKVIYLVMILLIAPA YSIRCIGVSNRDFVEGMSGGTWVDVVLEHGGCVTVMAQDKPAVDI ELVTTTVSNMAEVRSYCYEASISDMASDSRCPTQGEAYLDKQSDTQ YVCKRTLVDRGWGNGCGLFGKGSLVTCAKFACSKKMTGKSIQPEN LEYRIMLSVHGSQHSGMIVNDTGHETDENRAKVEITPNSPRAEATL GGFGSLGLDCEPRTGLDFSDLYYLTMNNKHWLVHKEWFHDIPLPW HAGADTGTPHWNNKEALVEFKDAHAKRQTVVVLGSQEGAVHTAL AGALEAEMDGAKGRLSSGHLKCRLKMDKLRLKGVSYSLCTAAFTF TKIPAETLHGTVTVEVQYAGTDGPCKVPAQMAVDMQTLTPVGRLI TANPVITESTENSKMMLELDPPFGDSYIVIGVGEKKITHHWHRSGST IGKAFEATVRGAKRMAVLGDTAWDFGSVGGALNSLGKGIHQIFGA AFKSLFGGMSWFSQILIGTLLVWLGLNTKNGSISLTCLALGGVLIFL STAVSA HuIgG_(k) signal METPAQLLFLLLLWLPDTTGVEVTRRGSAYYMYLDRSDAGEAISFP 206 peptide_prME #1 TTLGMNKCYIQIMDLGHMCDATMSYECPMLDEGVEPDDVDCWCN (Brazil_isolate_ TTSTWVVYGTCHHKKGEARRSRRAVTLPSHSTRKLQTRSQTWLES ZikaSPH2015) REYTKHLIRVENWIFRNPGFALAAAAIAWLLGSSTSQKVIYLVMILL IAPAYSIRCIGVSNRDFVEGMSGGTWVDVVLEHGGCVTVMAQDKP AVDIELVTTTVSNMAEVRSYCYEASISDMASDSRCPTQGEAYLDKQ SDTQYVCKRTLVDRGWGNGCGLFGKGSLVTCAKFACSKKMTGKSI QPENLEYRIMLSVHGSQHSGMIVNDTGHETDENRAKVEITPNSPRA EATLGGFGSLGLDCEPRTGLDFSDLYYLTMNNKHWLVHKEWFHDI PLPWHAGADTGTPHWNNKEALVEFKDAHAKRQTVVVLGSQEGAV HTALAGALEAEMDGAKGRLSSGHLKCRLKMDKLRLKGVSYSLCT AAFTFTKIPAETLHGTVTVEVQYAGTDGPCKVPAQMAVDMQTLTP VGRLITANPVITESTENSKMMLELDPPFGDSYIVIGVGEKKITHHWH RSGSTIGKAFEATVRGAKRMAVLGDTAWDFGSVGGALNSLGKGIH QIFGAAFKSLFGGMSWFSQILIGTLLVWLGLNTKNGSISLTCLALGG VLIFLSTAVSA HuIgG_(k) signal METPAQLLFLLLLWLPDTTGTRRGSAYYMYLDRSDAGEAISFPTTL 207 peptide_prME #2 GMNKCYIQIMDLGHMCDATMSYECPMLDEGVEPDDVDCWCNTTS (Brazil_isolate_ TWVVYGTCHHKKGEARRSRRAVTLPSHSTRKLQTRSQTWLESREY ZikaSPH2015) TKHLIRVENWIFRNPGFALAAAAIAWLLGSSTSQKVIYLVMILLIAP AYSIRCIGVSNRDFVEGMSGGTWVDVVLEHGGCVTVMAQDKPAV DIELVTTTVSNMAEVRSYCYEASISDMASDSRCPTQGEAYLDKQSD TQYVCKRTLVDRGWGNGCGLFGKGSLVTCAKFACSKKMTGKSIQP ENLEYRIMLSVHGSQHSGMIVNDTGHETDENRAKVEITPNSPRAEA TLGGFGSLGLDCEPRTGLDFSDLYYLTMNNKHWLVHKEWFHDIPL PWHAGADTGTPHWNNKEALVEFKDAHAKRQTVVVLGSQEGAVH TALAGALEAEMDGAKGRLSSGHLKCRLKMDKLRLKGVSYSLCTA AFTFTKIPAETLHGTVTVEVQYAGTDGPCKVPAQMAVDMQTLTPV GRLITANPVITESTENSKMMLELDPPFGDSYIVIGVGEKKITHHWHR SGSTIGKAFEATVRGAKRMAVLGDTAWDFGSVGGALNSLGKGIHQ IFGAAFKSLFGGMSWFSQILIGTLLVWLGLNTKNGSISLTCLALGGV LIFLSTAVSA HuIgG_(k) signal peptide_E METPAQLLFLLLLWLPDTTGIRCIGVSNRDFVEGMSGGTWVDVVLE 208 (Brazil_isolate_ HGGCVTVMAQDKPAVDIELVTTTVSNMAEVRSYCYEASISDMASD ZikaSPH2015) SRCPTQGEAYLDKQSDTQYVCKRTLVDRGWGNGCGLFGKGSLVT CAKFACSKKMTGKSIQPENLEYRIMLSVHGSQHSGMIVNDTGHETD ENRAKVEITPNSPRAEATLGGFGSLGLDCEPRTGLDFSDLYYLTMN NKHWLVHKEWFHDIPLPWHAGADTGTPHWNNKEALVEFKDAHA KRQTVVVLGSQEGAVHTALAGALEAEMDGAKGRLSSGHLKCRLK MDKLRLKGVSYSLCTAAFTFTKIPAETLHGTVTVEVQYAGTDGPCK VPAQMAVDMQTLTPVGRLITANPVITESTENSKMMLELDPPFGDSY IVIGVGEKKITHHWHRSGSTIGKAFEATVRGAKRMAVLGDTAWDF GSVGGALNSLGKGIHQIFGAAFKSLFGGMSWFSQILIGTLLVWLGL NTKNGSISLTCLALGGVLIFLSTAVSA IgE HC signal MDWTWILFLVAAATRVHSTRRGSAYYMYLDRSDAGEAISFPTTLG 209 peptide_prM-E #2 MNKCYIQIMDLGHMCDATMSYECPMLDEGVEPDDVDCWCNTTST (ACD75819_Micronesia) WVVYGTCHHKKGEARRSRRAVTLPSHSTRKLQTRSQTWLESREYT KHLIRVENWIFRNPGFALAAAAIAWLLGSSTSQKVIYLVMILLIAPA YSIRCIGVSNRDFVEGMSGGTWVDVVLEHGGCVTVMAQDKPAVDI ELVTTTVSNMAEVRSYCYEASISDMASDSRCPTQGEAYLDKQSDTQ YVCKRTLVDRGWGNGCGLFGKGSLVTCAKFACSKKMTGKSIQPEN LEYRIMLSVHGSQHSGMIVNDTGHETDENRAKVEITPNSPRAEATL GGFGSLGLDCEPRTGLDFSDLYYLTMNNKHWLVHKEWFHDIPLPW HAGADTGTPHWNNKEALVEFKDAHAKRQTVVVLGSQEGAVHTAL AGALEAEMDGAKGRLSSGHLKCRLKMDKLRLKGVSYSLCTAAFTF TKIPAETLHGTVTVEVQYAGTDGPCKVPAQMAVDMQTLTPVGRLI TANPVITESTENSKMMLELDPPFGDSYIVIGVGEKKITHHWHRSGST IGKAFEATVRGAKRMAVLGDTAWDFGSVGGALNSLGKGIHQIFGA AFKSLFGGMSWFSQILIGTLLVWLGLNTKNGSISLTCLALGGVLIFL STAVSA HuIgG_(k) signal METPAQLLFLLLLWLPDTTGVEVTRRGSAYYMYLDRSDAGEAISFP 210 peptide_prME #1, TTLGMNKCYIQIMDLGHMCDATMSYECPMLDEGVEPDDVDCWCN (ACD75819_Micronesia) TTSTWVVYGTCHHKKGEARRSRRAVTLPSHSTRKLQTRSQTWLES REYTKHLIRVENWIFRNPGFALAAAAIAWLLGSSTSQKVIYLVMILL IAPAYSIRCIGVSNRDFVEGMSGGTWVDVVLEHGGCVTVMAQDKP AVDIELVTTTVSNMAEVRSYCYEASISDMASDSRCPTQGEAYLDKQ SDTQYVCKRTLVDRGWGNGCGLFGKGSLVTCAKFACSKKMTGKSI QPENLEYRIMLSVHGSQHSGMIVNDTGHETDENRAKVEITPNSPRA EATLGGFGSLGLDCEPRTGLDFSDLYYLTMNNKHWLVHKEWFHDI PLPWHAGADTGTPHWNNKEALVEFKDAHAKRQTVVVLGSQEGAV HTALAGALEAEMDGAKGRLSSGHLKCRLKMDKLRLKGVSYSLCT AAFTFTKIPAETLHGTVTVEVQYAGTDGPCKVPAQMAVDMQTLTP VGRLITANPVITESTENSKMMLELDPPFGDSYIVIGVGEKKITHHWH RSGSTIGKAFEATVRGAKRMAVLGDTAWDFGSVGGALNSLGKGIH QIFGAAFKSLFGGMSWFSQILIGTLLVWLGLNTKNGSISLTCLALGG VLIFLSTAVSA HuIgG_(k) signal METPAQLLFLLLLWLPDTTGTRRGSAYYMYLDRSDAGEAISFPTTL 211 peptide_prME #2, GMNKCYIQIMDLGHMCDATMSYECPMLDEGVEPDDVDCWCNTTS (ACD75819_Micronesia) TWVVYGTCHHKKGEARRSRRAVTLPSHSTRKLQTRSQTWLESREY TKHLIRVENWIFRNPGFALAAAAIAWLLGSSTSQKVIYLVMILLIAP AYSIRCIGVSNRDFVEGMSGGTWVDVVLEHGGCVTVMAQDKPAV DIELVTTTVSNMAEVRSYCYEASISDMASDSRCPTQGEAYLDKQSD TQYVCKRTLVDRGWGNGCGLFGKGSLVTCAKFACSKKMTGKSIQP ENLEYRIMLSVHGSQHSGMIVNDTGHETDENRAKVEITPNSPRAEA TLGGFGSLGLDCEPRTGLDFSDLYYLTMNNKHWLVHKEWFHDIPL PWHAGADTGTPHWNNKEALVEFKDAHAKRQTVVVLGSQEGAVH TALAGALEAEMDGAKGRLSSGHLKCRLKMDKLRLKGVSYSLCTA AFTFTKIPAETLHGTVTVEVQYAGTDGPCKVPAQMAVDMQTLTPV GRLITANPVITESTENSKMMLELDPPFGDSYIVIGVGEKKITHHWHR SGSTIGKAFEATVRGAKRMAVLGDTAWDFGSVGGALNSLGKGIHQ IFGAAFKSLFGGMSWFSQILIGTLLVWLGLNTKNGSISLTCLALGGV LIFLSTAVSA HuIgG_(k) signal peptide_E, METPAQLLFLLLLWLPDTTGIRCIGVSNRDFVEGMSGGTWVDVVLE 212 (ACD75819_Micronesia) HGGCVTVMAQDKPAVDIELVTTTVSNMAEVRSYCYEASISDMASD SRCPTQGEAYLDKQSDTQYVCKRTLVDRGWGNGCGLFGKGSLVT CAKFACSKKMTGKSIQPENLEYRIMLSVHGSQHSGMIVNDTGHETD ENRAKVEITPNSPRAEATLGGFGSLGLDCEPRTGLDFSDLYYLTMN NKHWLVHKEWFHDIPLPWHAGADTGTPHWNNKEALVEFKDAHA KRQTVVVLGSQEGAVHTALAGALEAEMDGAKGRLSSGHLKCRLK MDKLRLKGVSYSLCTAAFTFTKIPAETLHGTVTVEVQYAGTDGPCK VPAQMAVDMQTLTPVGRLITANPVITESTENSKMMLELDPPFGDSY IVIGVGEKKITHHWHRSGSTIGKAFEATVRGAKRMAVLGDTAWDF GSVGGALNSLGKGIHQIFGAAFKSLFGGMSWFSQILIGTLLVWLGL NTKNGSISLTCLALGGVLIFLSTAVSA Zika_RIO-U1_JEVsp AEVTRRGSAYYMYLDRNDAGEAISFPTTLGMNKCYIQIMDLGHMC 213 Zika PRME Strain DATMSYECPMLDEGVEPDDVDCWCNTTSTWVVYGTCHHKKGEA ascension id: ANG09399 RRSRRAVTLPSHSTRKLQTRSQTWLESREYTKHLIRVENWIFRNPGF ALAAAAIAWLLGSSTSQKVIYLVMILLIAPAYSIRCIGVSNRDFVEG MSGGTWVDVVLEHGGCVTVMAQDKPTVDIELVTTTVSNMAEVRS YCYEASISDMASDSRCPTQGEAYLDKQSDTQYVCKRTLVDRGWGN GCGLFGKGSLVTCAKFACSKKMTGKSIQPENLEYRIMLSVHGSQHS GMIVNDTGHETDENRAKVEITPNSPRAEATLGGFGSLGLDCEPRTG LDFSDLYYLTMNNKHWLVHKEWFHDIPLPWHAGADTGTPHWNN KEALVEFKDAHAKRQTVVVLGSQEGAVHTALAGALEAEMDGAKG RLSSGHLKCRLKMDKLRLKGVSYSLCTAAFTFTKIPAETLHGTVTV EVQYAGTDGPCKVPAQMAVDMQTLTPVGRLITANPVITESTENSK MMLELDPPFGDSYIVIGVGEKKITHHWHRSGSTIGKAFEATVRGAK RMAVLGDTAWDFGSVGGALNSLGKGIHQIFGAAFKSLFGGMSWFS QILIGTLLMWLGLNTKNGSISLMCLALGGVLIFLSTAVSA Zika_RIO-U1_JEVsp MLGSNSGQRVVFTILLLLVAPAYSAEVTRRGSAYYMYLDRNDAGE 214 Zika PRME Strain AISFPTTLGMNKCYIQIMDLGHMCDATMSYECPMLDEGVEPDDVD ascension id: ANG09399 CWCNTTSTWVVYGTCHHKKGEARRSRRAVTLPSHSTRKLQTRSQT with JEVPRM signal WLESREYTKHLIRVENWIFRNPGFALAAAAIAWLLGSSTSQKVIYL sequence VMILLIAPAYSIRCIGVSNRDFVEGMSGGTWVDVVLEHGGCVTVM AQDKPTVDIELVTTTVSNMAEVRSYCYEASISDMASDSRCPTQGEA YLDKQSDTQYVCKRTLVDRGWGNGCGLFGKGSLVTCAKFACSKK MTGKSIQPENLEYRIMLSVHGSQHSGMIVNDTGHETDENRAKVEIT PNSPRAEATLGGFGSLGLDCEPRTGLDFSDLYYLTMNNKHWLVHK EWFHDIPLPWHAGADTGTPHWNNKEALVEFKDAHAKRQTVVVLG SQEGAVHTALAGALEAEMDGAKGRLSSGHLKCRLKMDKLRLKGV SYSLCTAAFTFTKIPAETLHGTVTVEVQYAGTDGPCKVPAQMAVD MQTLTPVGRLITANPVITESTENSKMMLELDPPFGDSYIVIGVGEKKI THHWHRSGSTIGKAFEATVRGAKRMAVLGDTAWDFGSVGGALNS LGKGIHQIFGAAFKSLFGGMSWFSQILIGTLLMWLGLNTKNGSISLM CLALGGVLIFLSTAVSA Zika_ RIO-U1¬_VSVgSp EVTRRGSAYYMYLDRNDAGEAISFPTTLGMNKCYIQIMDLGHMCD 215 Zika PRME Strain ATMSYECPMLDEGVEPDDVDCWCNTTSTWVVYGTCHHKKGEAR ascension id: ANG09399 RSRRAVTLPSHSTRKLQTRSQTWLESREYTKHLIRVENWIFRNPGFA LAAAAIAWLLGSSTSQKVIYLVMILLIAPAYSIRCIGVSNRDFVEGM SGGTWVDVVLEHGGCVTVMAQDKPTVDIELVTTTVSNMAEVRSY CYEASISDMASDSRCPTQGEAYLDKQSDTQYVCKRTLVDRGWGNG CGLFGKGSLVTCAKFACSKKMTGKSIQPENLEYRIMLSVHGSQHSG MIVNDTGHETDENRAKVEITPNSPRAEATLGGFGSLGLDCEPRTGL DFSDLYYLTMNNKHWLVHKEWFHDIPLPWHAGADTGTPHWNNK EALVEFKDAHAKRQTVVVLGSQEGAVHTALAGALEAEMDGAKGR LSSGHLKCRLKMDKLRLKGVSYSLCTAAFTFTKIPAETLHGTVTVE VQYAGTDGPCKVPAQMAVDMQTLTPVGRLITANPVITESTENSKM MLELDPPFGDSYIVIGVGEKKITHHWHRSGSTIGKAFEATVRGAKR MAVLGDTAWDFGSVGGALNSLGKGIHQIFGAAFKSLFGGMSWFSQ ILIGTLLMWLGLNTKNGSISLMCLALGGVLIFLSTAVSA Zika_ RIO-U1¬_VSVgSp MKCLLYLAFLFIGVNCAEVTRRGSAYYMYLDRNDAGEAISFPTTLG 216 Zika PRME Strain MNKCYIQIMDLGHMCDATMSYECPMLDEGVEPDDVDCWCNTTST ascension id: ANG09399 WVVYGTCHHKKGEARRSRRAVTLPSHSTRKLQTRSQTWLESREYT with VSVg protein signal KHLIRVENWIFRNPGFALAAAAIAWLLGSSTSQKVIYLVMILLIAPA sequence YSIRCIGVSNRDFVEGMSGGTWVDVVLEHGGCVTVMAQDKPTVDI ELVTTTVSNMAEVRSYCYEASISDMASDSRCPTQGEAYLDKQSDTQ YVCKRTLVDRGWGNGCGLFGKGSLVTCAKFACSKKMTGKSIQPEN LEYRIMLSVHGSQHSGMIVNDTGHETDENRAKVEITPNSPRAEATL GGFGSLGLDCEPRTGLDFSDLYYLTMNNKHWLVHKEWFHDIPLPW HAGADTGTPHWNNKEALVEFKDAHAKRQTVVVLGSQEGAVHTAL AGALEAEMDGAKGRLSSGHLKCRLKMDKLRLKGVSYSLCTAAFTF TKIPAETLHGTVTVEVQYAGTDGPCKVPAQMAVDMQTLTPVGRLI TANPVITESTENSKMMLELDPPFGDSYIVIGVGEKKITHHWHRSGST IGKAFEATVRGAKRMAVLGDTAWDFGSVGGALNSLGKGIHQIFGA AFKSLFGGMSWFSQILIGTLLMWLGLNTKNGSISLMCLALGGVLIFL STAVSA ZIKA_PRME_DSP_N154 VEVTRRGSAYYMYLDRSDAGEAISFPTTLGMNKCYIQIMDLGHMC 217 A (glycosylation mutant) DATMSYECPMLDEGVEPDDVDCWCNTTSTWVVYGTCHHKKGEA Zika PRME Strain RRSRRAVTLPSHSTRKLQTRSQTWLESREYTKHLIRVENWIFRNPGF ascension id: ACD75819 ALAAAAIAWLLGSSTSQKVIYLVMILLIAPAYSIRCIGVSNRDFVEG MSGGTWVDVVLEHGGCVTVMAQDKPAVDIELVTTTVSNMAEVRS YCYEASISDMASDSRCPTQGEAYLDKQSDTQYVCKRTLVDRGWGN GCGLFGKGSLVTCAKFACSKKMTGKSIQPENLEYRIMLSVHGSQHS GMIVADTGHETDENRAKVEITPNSPRAEATLGGFGSLGLDCEPRTG LDFSDLYYLTMNNKHWLVHKEWFHDIPLPWHAGADTGTPHWNN KEALVEFKDAHAKRQTVVVLGSQEGAVHTALAGALEAEMDGAKG RLSSGHLKCRLKMDKLRLKGVSYSLCTAAFTFTKIPAETLHGTVTV EVQYAGTDGPCKVPAQMAVDMQTLTPVGRLITANPVITESTENSK MMLELDPPFGDSYIVIGVGEKKITHHWHRSGSTIGKAFEATVRGAK RMAVLGDTAWDFGSVGGALNSLGKGIHQIFGAAFKSLFGGMSWFS QILIGTLLVWLGLNTKNGSISLTCLALGGVLIFLSTAVSA ZIKA_PRME_DSP_N154 MDWTWILFLVAAATRVHSVEVTRRGSAYYMYLDRSDAGEAISFPT 218 A (glycosylation mutant TLGMNKCYIQIMDLGHMCDATMSYECPMLDEGVEPDDVDCWCNT with signal peptide) TSTWVVYGTCHHKKGEARRSRRAVTLPSHSTRKLQTRSQTWLESR Zika PRME Strain EYTKHLIRVENWIFRNPGFALAAAAIAWLLGSSTSQKVIYLVMILLI ascension id: ACD75819 APAYSIRCIGVSNRDFVEGMSGGTWVDVVLEHGGCVTVMAQDKP with IgE signal peptide AVDIELVTTTVSNMAEVRSYCYEASISDMASDSRCPTQGEAYLDKQ SDTQYVCKRTLVDRGWGNGCGLFGKGSLVTCAKFACSKKMTGKSI QPENLEYRIMLSVHGSQHSGMIVADTGHETDENRAKVEITPNSPRA EATLGGFGSLGLDCEPRTGLDFSDLYYLTMNNKHWLVHKEWFHDI PLPWHAGADTGTPHWNNKEALVEFKDAHAKRQTVVVLGSQEGAV HTALAGALEAEMDGAKGRLSSGHLKCRLKMDKLRLKGVSYSLCT AAFTFTKIPAETLHGTVTVEVQYAGTDGPCKVPAQMAVDMQTLTP VGRLITANPVITESTENSKMMLELDPPFGDSYIVIGVGEKKITHHWH RSGSTIGKAFEATVRGAKRMAVLGDTAWDFGSVGGALNSLGKGIH QIFGAAFKSLFGGMSWFSQILIGTLLVWLGLNTKNGSISLTCLALGG VLIFLSTAVSA Zika_JEVsp_prME_V2 MWLVSLAIVTACAGAAEVTRRGSAYYMYLDRNDAGEAISFPTTLG 219 (signal peptide MNKCYIQIMDLGHMCDATMSYECPMLDEGVEPDDVDCWCNTTST underlined) WVVYGTCHHKKGEARRSRRAVTLPSHSTRKLQTRSQTWLESREYT KHLIRVENWIFRNPGFALAAAAIAWLLGSSTSQKVIYLVMILLIAPA YSIRCIGVSNRDFVEGMSGGTWVDVVLEHGGCVTVMAQDKPTVDI ELVTTTVSNMAEVRSYCYEASISDMASDSRCPTQGEAYLDKQSDTQ YVCKRTLVDRGWGNGCGLFGKGSLVTCAKFACSKKMTGKSIQPEN LEYRIMLSVHGSQHSGMIVNDTGHETDENRAKVEITPNSPRAEATL GGFGSLGLDCEPRTGLDFSDLYYLTMNNKHWLVHKEWFHDIPLPW HAGADTGTPHWNNKEALVEFKDAHAKRQTVVVLGSQEGAVHTAL AGALEAEMDGAKGRLSSGHLKCRLKMDKLRLKGVSYSLCTAAFTF TKIPAETLHGTVTVEVQYAGTDGPCKVPAQMAVDMQTLTPVGRLI TANPVITESTENSKMMLELDPPFGDSYIVIGVGEKKITHHWHRSGST IGKAFEATVRGAKRMAVLGDTAWDFGSVGGALNSLGKGIHQIFGA AFKSLFGGMSWFSQILIGTLLMWLGLNTKNGSISLMCLALGGVLIFL STAVSA Zika_JEVsp_prME_V2 AEVTRRGSAYYMYLDRNDAGEAISFPTTLGMNKCYIQIMDLGHMC 220 (no signal peptide) DATMSYECPMLDEGVEPDDVDCWCNTTSTWVVYGTCHHKKGEA RRSRRAVTLPSHSTRKLQTRSQTWLESREYTKHLIRVENWIFRNPGF ALAAAAIAWLLGSSTSQKVIYLVMILLIAPAYSIRCIGVSNRDFVEG MSGGTWVDVVLEHGGCVTVMAQDKPTVDIELVTTTVSNMAEVRS YCYEASISDMASDSRCPTQGEAYLDKQSDTQYVCKRTLVDRGWGN GCGLFGKGSLVTCAKFACSKKMTGKSIQPENLEYRIMLSVHGSQHS GMIVNDTGHETDENRAKVEITPNSPRAEATLGGFGSLGLDCEPRTG LDFSDLYYLTMNNKHWLVHKEWFHDIPLPWHAGADTGTPHWNN KEALVEFKDAHAKRQTVVVLGSQEGAVHTALAGALEAEMDGAKG RLSSGHLKCRLKMDKLRLKGVSYSLCTAAFTFTKIPAETLHGTVTV EVQYAGTDGPCKVPAQMAVDMQTLTPVGRLITANPVITESTENSK MMLELDPPFGDSYIVIGVGEKKITHHWHRSGSTIGKAFEATVRGAK RMAVLGDTAWDFGSVGGALNSLGKGIHQIFGAAFKSLFGGMSWFS QILIGTLLMWLGLNTKNGSISLMCLALGGVLIFLSTAVSA Zika_JEVPRSp_prME_4 MWLVSLAIVTACAGAAEVTRRGSAYYMYLDRNDAGEAISFPTTLG 221 Mut_V2 MNKCYIQIMDLGHMCDATMSYECPMLDEGVEPDDVDCWCNTTST (signal peptide WVVYGTCHHKKGEARRSRRAVTLPSHSTRKLQTRSQTWLESREYT underlined) KHLIRVENWIFRNPGFALAAAAIAWLLGSSTSQKVIYLVMILLIAPA YSIRCIGVSNRDFVEGMSGGTWVDVVLEHGGCVTVMAQDKPTVDI ELVTTTVSNMAEVRSYCYEASISDMASDSRCPREGEAYLDKQSDTQ YVCKRTLVDRGRGNGCGRFGKGSLVTCAKFACSKKMTGKSIQPEN LEYRIMLSVHGSQHSGMIVNDTGHETDENRAKVEITPNSPRAEATL GGFGSLGLDCEPRTGLDFSDLYYLTMNNKHWLVHKEWFHDIPLPW HAGADTGTPHWNNKEALVEFKDAHAKRQTVVVLGSQEGAVHTAL AGALEAEMDGAKGRLSSGHLKCRLKMDKLRLKGVSYSLCTAAFTF TKIPAETLHGTVTVEVQYAGTDGPCKVPAQMAVDMQTLTPVGRLI TANPVITESTENSKMMLELDPPFGDSYIVIGVGEKKITHHWHRSGST IGKAFEATVRGAKRMAVLGDTAWDFGSVGGALNSLGKGIHQIFGA AFKSLFGGMSWFSQILIGTLLMWLGLNTKNGSISLMCLALGGVLIFL STAVSA Zika_JEVPRSp_prME_4 AEVTRRGSAYYMYLDRNDAGEAISFPTTLGMNKCYIQIMDLGHMC 222 Mut_V2 DATMSYECPMLDEGVEPDDVDCWCNTTSTWVVYGTCHHKKGEA (no signal peptide) RRSRRAVTLPSHSTRKLQTRSQTWLESREYTKHLIRVENWIFRNPGF ALAAAAIAWLLGSSTSQKVIYLVMILLIAPAYSIRCIGVSNRDFVEG MSGGTWVDVVLEHGGCVTVMAQDKPTVDIELVTTTVSNMAEVRS YCYEASISDMASDSRCPREGEAYLDKQSDTQYVCKRTLVDRGRGN GCGRFGKGSLVTCAKFACSKKMTGKSIQPENLEYRIMLSVHGSQHS GMIVNDTGHETDENRAKVEITPNSPRAEATLGGFGSLGLDCEPRTG LDFSDLYYLTMNNKHWLVHKEWFHDIPLPWHAGADTGTPHWNN KEALVEFKDAHAKRQTVVVLGSQEGAVHTALAGALEAEMDGAKG RLSSGHLKCRLKMDKLRLKGVSYSLCTAAFTFTKIPAETLHGTVTV EVQYAGTDGPCKVPAQMAVDMQTLTPVGRLITANPVITESTENSK MMLELDPPFGDSYIVIGVGEKKITHHWHRSGSTIGKAFEATVRGAK RMAVLGDTAWDFGSVGGALNSLGKGIHQIFGAAFKSLFGGMSWFS QILIGTLLMWLGLNTKNGSISLMCLALGGVLIFLSTAVSA

Underlined sequence corresponds to a signal peptide, which may be omitted from each sequence. Thus, any RNA vaccine provided herein may encode an antigen represented by a sequence of Table 26, with or without the underlined signal peptide.

TABLE 27 ZIKV NCBI Accession Numbers (Amino Acid Sequences) Name GenBank Accession polyprotein [Zika virus] YP_002790881.1 polyprotein [Zika virus] BAP47441.1 polyprotein [Zika virus] AEN75263.1 polyprotein [Zika virus] AHL43504.1 polyprotein [Zika virus] AEN75266.1 polyprotein [Zika virus] AHF49784.1 polyprotein [Zika virus] AHF49783.1 polyprotein [Zika virus] AHF49785.1 polyprotein [Zika virus] ABI54475.1 polyprotein [Zika virus] AHL43501.1 polyprotein [Zika virus] AHL43500.1 polyprotein [Zika virus] AHL43502.1 polyprotein [Zika virus] AEN75265.1 polyprotein [Zika virus] AHL43503.1 polyprotein [Zika virus] AEN75264.1 polyprotein [Zika virus] AHZ13508.1 polyprotein [Zika virus] ACD75819.1 polyprotein [Zika virus] AFD30972.1 polyprotein [Zika virus] AAK91609.1 envelope protein [Zika virus] AHL43462.1 envelope protein [Zika virus] AHL43464.1 envelope protein [Zika virus] AHL43461.1 envelope protein [Zika virus] AHL43460.1 envelope protein [Zika virus] AHL43463.1 envelope protein [Zika virus] AHL43444.1 envelope protein [Zika virus] AHL43451.1 envelope protein [Zika virus] AHL43437.1 envelope protein [Zika virus] AHL43455.1 envelope protein [Zika virus] AHL43448.1 envelope protein [Zika virus] AHL43439.1 envelope protein [Zika virus] AHL43468.1 E protein [Zika virus] AIC06934.1 envelope protein [Zika virus] AHL43450.1 envelope protein [Zika virus] AHL43442.1 envelope protein [Zika virus] AHL43458.1 envelope glycoprotein [Zika virus] AHL16749.1 envelope protein [Zika virus] AHL43453.1 envelope protein [Zika virus] AHL43443.1 envelope protein [Zika virus] AHL43438.1 envelope protein [Zika virus] AHL43441.1 envelope protein [Zika virus] AHL43457.1 envelope protein [Zika virus] AAK91609.1 polyprotein [Zika virus] AHL43505.1

TABLE 28 DENV polynucleotide sequences SEQ ID Name Sequence NO: DEN-1 AGTTGTTAGTCTACGTGGACCGACAAGAACAGTTTCGAATCGGAA 223 (NC_001477.1) GCTTGCTTAACGTAGTTCTAACAGTTTTTTATTAGAGAGCAGATCT CTGATGAACAACCAACGGAAAAAGACGGGTCGACCGTCTTTCAAT ATGCTGAAACGCGCGAGAAACCGCGTGTCAACTGTTTCACAGTTG GCGAAGAGATTCTCAAAAGGATTGCTTTCAGGCCAAGGACCCATG AAATTGGTGATGGCTTTTATAGCATTCCTAAGATTTCTAGCCATAC CTCCAACAGCAGGAATTTTGGCTAGATGGGGCTCATTCAAGAAGA ATGGAGCGATCAAAGTGTTACGGGGTTTCAAGAAAGAAATCTCAA ACATGTTGAACATAATGAACAGGAGGAAAAGATCTGTGACCATGC TCCTCATGCTGCTGCCCACAGCCCTGGCGTTCCATCTGACCACCCG AGGGGGAGAGCCGCACATGATAGTTAGCAAGCAGGAAAGAGGAA AATCACTTTTGTTTAAGACCTCTGCAGGTGTCAACATGTGCACCCT TATTGCAATGGATTTGGGAGAGTTATGTGAGGACACAATGACCTA CAAATGCCCCCGGATCACTGAGACGGAACCAGATGACGTTGACTG TTGGTGCAATGCCACGGAGACATGGGTGACCTATGGAACATGTTC TCAAACTGGTGAACACCGACGAGACAAACGTTCCGTCGCACTGGC ACCACACGTAGGGCTTGGTCTAGAAACAAGAACCGAAACGTGGAT GTCCTCTGAAGGCGCTTGGAAACAAATACAAAAAGTGGAGACCTG GGCTCTGAGACACCCAGGATTCACGGTGATAGCCCTTTTTCTAGCA CATGCCATAGGAACATCCATCACCCAGAAAGGGATCATTTTTATTT TGCTGATGCTGGTAACTCCATCCATGGCCATGCGGTGCGTGGGAAT AGGCAACAGAGACTTCGTGGAAGGACTGTCAGGAGCTACGTGGGT GGATGTGGTACTGGAGCATGGAAGTTGCGTCACTACCATGGCAAA AGACAAACCAACACTGGACATTGAACTCTTGAAGACGGAGGTCAC AAACCCTGCCGTCCTGCGCAAACTGTGCATTGAAGCTAAAATATC AAACACCACCACCGATTCGAGATGTCCAACACAAGGAGAAGCCAC GCTGGTGGAAGAACAGGACACGAACTTTGTGTGTCGACGAACGTT CGTGGACAGAGGCTGGGGCAATGGTTGTGGGCTATTCGGAAAAGG TAGCTTAATAACGTGTGCTAAGTTTAAGTGTGTGACAAAACTGGA AGGAAAGATAGTCCAATATGAAAACTTAAAATATTCAGTGATAGT CACCGTACACACTGGAGACCAGCACCAAGTTGGAAATGAGACCAC AGAACATGGAACAACTGCAACCATAACACCTCAAGCTCCCACGTC GGAAATACAGCTGACAGACTACGGAGCTCTAACATTGGATTGTTC ACCTAGAACAGGGCTAGACTTTAATGAGATGGTGTTGTTGACAAT GAAAAAAAAATCATGGCTCGTCCACAAACAATGGTTTCTAGACTT ACCACTGCCTTGGACCTCGGGGGCTTCAACATCCCAAGAGACTTG GAATAGACAAGACTTGCTGGTCACATTTAAGACAGCTCATGCAAA AAAGCAGGAAGTAGTCGTACTAGGATCACAAGAAGGAGCAATGC ACACTGCGTTGACTGGAGCGACAGAAATCCAAACGTCTGGAACGA CAACAATTTTTGCAGGACACCTGAAATGCAGATTAAAAATGGATA AACTGATTTTAAAAGGGATGTCATATGTAATGTGCACAGGGTCATT CAAGTTAGAGAAGGAAGTGGCTGAGACCCAGCATGGAACTGTTCT AGTGCAGGTTAAATACGAAGGAACAGATGCACCATGCAAGATCCC CTTCTCGTCCCAAGATGAGAAGGGAGTAACCCAGAATGGGAGATT GATAACAGCCAACCCCATAGTCACTGACAAAGAAAAACCAGTCAA CATTGAAGCGGAGCCACCTTTTGGTGAGAGCTACATTGTGGTAGG AGCAGGTGAAAAAGCTTTGAAACTAAGCTGGTTCAAGAAGGGAA GCAGTATAGGGAAAATGTTTGAAGCAACTGCCCGTGGAGCACGAA GGATGGCCATCCTGGGAGACACTGCATGGGACTTCGGTTCTATAG GAGGGGTGTTCACGTCTGTGGGAAAACTGATACACCAGATTTTTG GGACTGCGTATGGAGTTTTGTTCAGCGGTGTTTCTTGGACCATGAA GATAGGAATAGGGATTCTGCTGACATGGCTAGGATTAAACTCAAG GAGCACGTCCCTTTCAATGACGTGTATCGCAGTTGGCATGGTCACA CTGTACCTAGGAGTCATGGTTCAGGCGGACTCGGGATGTGTAATC AACTGGAAAGGCAGAGAACTCAAATGTGGAAGCGGCATTTTTGTC ACCAATGAAGTCCACACCTGGACAGAGCAATATAAATTCCAGGCC GACTCCCCTAAGAGACTATCAGCGGCCATTGGGAAGGCATGGGAG GAGGGTGTGTGTGGAATTCGATCAGCCACTCGTCTCGAGAACATC ATGTGGAAGCAAATATCAAATGAATTAAACCACATCTTACTTGAA AATGACATGAAATTTACAGTGGTCGTAGGAGACGTTAGTGGAATC TTGGCCCAAGGAAAGAAAATGATTAGGCCACAACCCATGGAACAC AAATACTCGTGGAAAAGCTGGGGAAAAGCCAAAATCATAGGAGC AGATGTACAGAATACCACCTTCATCATCGACGGCCCAAACACCCC AGAATGCCCTGATAACCAAAGAGCATGGAACATTTGGGAAGTTGA AGACTATGGATTTGGAATTTTCACGACAAACATATGGTTGAAATTG CGTGACTCCTACACTCAAGTGTGTGACCACCGGCTAATGTCAGCTG CCATCAAGGATAGCAAAGCAGTCCATGCTGACATGGGGTACTGGA TAGAAAGTGAAAAGAACGAGACTTGGAAGTTGGCAAGAGCCTCCT TCATAGAAGTTAAGACATGCATCTGGCCAAAATCCCACACTCTAT GGAGCAATGGAGTCCTGGAAAGTGAGATGATAATCCCAAAGATAT ATGGAGGACCAATATCTCAGCACAACTACAGACCAGGATATTTCA CACAAACAGCAGGGCCGTGGCACTTGGGCAAGTTAGAACTAGATT TTGATTTATGTGAAGGTACCACTGTTGTTGTGGATGAACATTGTGG AAATCGAGGACCATCTCTTAGAACCACAACAGTCACAGGAAAGAC AATCCATGAATGGTGCTGTAGATCTTGCACGTTACCCCCCCTACGT TTCAAAGGAGAAGACGGGTGCTGGTACGGCATGGAAATCAGACCA GTCAAGGAGAAGGAAGAGAACCTAGTTAAGTCAATGGTCTCTGCA GGGTCAGGAGAAGTGGACAGTTTTTCACTAGGACTGCTATGCATA TCAATAATGATCGAAGAGGTAATGAGATCCAGATGGAGCAGAAA AATGCTGATGACTGGAACATTGGCTGTGTTCCTCCTTCTCACAATG GGACAATTGACATGGAATGATCTGATCAGGCTATGTATCATGGTT GGAGCCAACGCTTCAGACAAGATGGGGATGGGAACAACGTACCTA GCTTTGATGGCCACTTTCAGAATGAGACCAATGTTCGCAGTCGGGC TACTGTTTCGCAGATTAACATCTAGAGAAGTTCTTCTTCTTACAGT TGGATTGAGTCTGGTGGCATCTGTAGAACTACCAAATTCCTTAGAG GAGCTAGGGGATGGACTTGCAATGGGCATCATGATGTTGAAATTA CTGACTGATTTTCAGTCACATCAGCTATGGGCTACCTTGCTGTCTTT AACATTTGTCAAAACAACTTTTTCATTGCACTATGCATGGAAGACA ATGGCTATGATACTGTCAATTGTATCTCTCTTCCCTTTATGCCTGTC CACGACTTCTCAAAAAACAACATGGCTTCCGGTGTTGCTGGGATCT CTTGGATGCAAACCACTAACCATGTTTCTTATAACAGAAAACAAA ATCTGGGGAAGGAAAAGCTGGCCTCTCAATGAAGGAATTATGGCT GTTGGAATAGTTAGCATTCTTCTAAGTTCACTTCTCAAGAATGATG TGCCACTAGCTGGCCCACTAATAGCTGGAGGCATGCTAATAGCAT GTTATGTCATATCTGGAAGCTCGGCCGATTTATCACTGGAGAAAGC GGCTGAGGTCTCCTGGGAAGAAGAAGCAGAACACTCTGGTGCCTC ACACAACATACTAGTGGAGGTCCAAGATGATGGAACCATGAAGAT AAAGGATGAAGAGAGAGATGACACACTCACCATTCTCCTCAAAGC AACTCTGCTAGCAATCTCAGGGGTATACCCAATGTCAATACCGGC GACCCTCTTTGTGTGGTATTTTTGGCAGAAAAAGAAACAGAGATC AGGAGTGCTATGGGACACACCCAGCCCTCCAGAAGTGGAAAGAGC AGTCCTTGATGATGGCATTTATAGAATTCTCCAAAGAGGATTGTTG GGCAGGTCTCAAGTAGGAGTAGGAGTTTTTCAAGAAGGCGTGTTC CACACAATGTGGCACGTCACCAGGGGAGCTGTCCTCATGTACCAA GGGAAGAGACTGGAACCAAGTTGGGCCAGTGTCAAAAAAGACTT GATCTCATATGGAGGAGGTTGGAGGTTTCAAGGATCCTGGAACGC GGGAGAAGAAGTGCAGGTGATTGCTGTTGAACCGGGGAAGAACC CCAAAAATGTACAGACAGCGCCGGGTACCTTCAAGACCCCTGAAG GCGAAGTTGGAGCCATAGCTCTAGACTTTAAACCCGGCACATCTG GATCTCCTATCGTGAACAGAGAGGGAAAAATAGTAGGTCTTTATG GAAATGGAGTGGTGACAACAAGTGGTACCTACGTCAGTGCCATAG CTCAAGCTAAAGCATCACAAGAAGGGCCTCTACCAGAGATTGAGG ACGAGGTGTTTAGGAAAAGAAACTTAACAATAATGGACCTACATC CAGGATCGGGAAAAACAAGAAGATACCTTCCAGCCATAGTCCGTG AGGCCATAAAAAGAAAGCTGCGCACGCTAGTCTTAGCTCCCACAA GAGTTGTCGCTTCTGAAATGGCAGAGGCGCTCAAGGGAATGCCAA TAAGGTATCAGACAACAGCAGTGAAGAGTGAACACACGGGAAAG GAGATAGTTGACCTTATGTGTCACGCCACTTTCACTATGCGTCTCC TGTCTCCTGTGAGAGTTCCCAATTATAATATGATTATCATGGATGA AGCACATTTTACCGATCCAGCCAGCATAGCAGCCAGAGGGTATAT CTCAACCCGAGTGGGTATGGGTGAAGCAGCTGCGATTTTCATGAC AGCCACTCCCCCCGGATCGGTGGAGGCCTTTCCACAGAGCAATGC AGTTATCCAAGATGAGGAAAGAGACATTCCTGAAAGATCATGGAA CTCAGGCTATGACTGGATCACTGATTTCCCAGGTAAAACAGTCTGG TTTGTTCCAAGCATCAAATCAGGAAATGACATTGCCAACTGTTTAA GAAAGAATGGGAAACGGGTGGTCCAATTGAGCAGAAAAACTTTTG ACACTGAGTACCAGAAAACAAAAAATAACGACTGGGACTATGTTG TCACAACAGACATATCCGAAATGGGAGCAAACTTCCGAGCCGACA GGGTAATAGACCCGAGGCGGTGCCTGAAACCGGTAATACTAAAAG ATGGCCCAGAGCGTGTCATTCTAGCCGGACCGATGCCAGTGACTG TGGCTAGCGCCGCCCAGAGGAGAGGAAGAATTGGAAGGAACCAA AATAAGGAAGGCGATCAGTATATTTACATGGGACAGCCTCTAAAC AATGATGAGGACCACGCCCATTGGACAGAAGCAAAAATGCTCCTT GACAACATAAACACACCAGAAGGGATTATCCCAGCCCTCTTTGAG CCGGAGAGAGAAAAGAGTGCAGCAATAGACGGGGAATACAGACT ACGGGGTGAAGCGAGGAAAACGTTCGTGGAGCTCATGAGAAGAG GAGATCTACCTGTCTGGCTATCCTACAAAGTTGCCTCAGAAGGCTT CCAGTACTCCGACAGAAGGTGGTGCTTTGATGGGGAAAGGAACAA CCAGGTGTTGGAGGAGAACATGGACGTGGAGATCTGGACAAAAG AAGGAGAAAGAAAGAAACTACGACCCCGCTGGCTGGATGCCAGA ACATACTCTGACCCACTGGCTCTGCGCGAATTCAAAGAGTTCGCA GCAGGAAGAAGAAGCGTCTCAGGTGACCTAATATTAGAAATAGGG AAACTTCCACAACATTTAACGCAAAGGGCCCAGAACGCCTTGGAC AATCTGGTTATGTTGCACAACTCTGAACAAGGAGGAAAAGCCTAT AGACACGCCATGGAAGAACTACCAGACACCATAGAAACGTTAATG CTCCTAGCTTTGATAGCTGTGCTGACTGGTGGAGTGACGTTGTTCT TCCTATCAGGAAGGGGTCTAGGAAAAACATCCATTGGCCTACTCT GCGTGATTGCCTCAAGTGCACTGTTATGGATGGCCAGTGTGGAAC CCCATTGGATAGCGGCCTCTATCATACTGGAGTTCTTTCTGATGGT GTTGCTTATTCCAGAGCCGGACAGACAGCGCACTCCACAAGACAA CCAGCTAGCATACGTGGTGATAGGTCTGTTATTCATGATATTGACA GTGGCAGCCAATGAGATGGGATTACTGGAAACCACAAAGAAGGA CCTGGGGATTGGTCATGCAGCTGCTGAAAACCACCATCATGCTGC AATGCTGGACGTAGACCTACATCCAGCTTCAGCCTGGACTCTCTAT GCAGTGGCCACAACAATTATCACTCCCATGATGAGACACACAATT GAAAACACAACGGCAAATATTTCCCTGACAGCTATTGCAAACCAG GCAGCTATATTGATGGGACTTGACAAGGGATGGCCAATATCAAAG ATGGACATAGGAGTTCCACTTCTCGCCTTGGGGTGCTATTCTCAGG TGAACCCGCTGACGCTGACAGCGGCGGTATTGATGCTAGTGGCTC ATTATGCCATAATTGGACCCGGACTGCAAGCAAAAGCTACTAGAG AAGCTCAAAAAAGGACAGCAGCCGGAATAATGAAAAACCCAACT GTCGACGGGATCGTTGCAATAGATTTGGACCCTGTGGTTTACGATG CAAAATTTGAAAAACAGCTAGGCCAAATAATGTTGTTGATACTTT GCACATCACAGATCCTCCTGATGCGGACCACATGGGCCTTGTGTG AATCCATCACACTAGCCACTGGACCTCTGACTACGCTTTGGGAGG GATCTCCAGGAAAATTCTGGAACACCACGATAGCGGTGTCCATGG CAAACATTTTTAGGGGAAGTTATCTAGCAGGAGCAGGTCTGGCCT TTTCATTAATGAAATCTCTAGGAGGAGGTAGGAGAGGCACGGGAG CCCAAGGGGAAACACTGGGAGAAAAATGGAAAAGACAGCTAAAC CAATTGAGCAAGTCAGAATTCAACACTTACAAAAGGAGTGGGATT ATAGAGGTGGATAGATCTGAAGCCAAAGAGGGGTTAAAAAGAGG AGAAACGACTAAACACGCAGTGTCGAGAGGAACGGCCAAACTGA GGTGGTTTGTGGAGAGGAACCTTGTGAAACCAGAAGGGAAAGTCA TAGACCTCGGTTGTGGAAGAGGTGGCTGGTCATATTATTGCGCTGG GCTGAAGAAAGTCACAGAAGTGAAAGGATACACGAAAGGAGGAC CTGGACATGAGGAACCAATCCCAATGGCAACCTATGGATGGAACC TAGTAAAGCTATACTCCGGGAAAGATGTATTCTTTACACCACCTGA GAAATGTGACACCCTCTTGTGTGATATTGGTGAGTCCTCTCCGAAC CCAACTATAGAAGAAGGAAGAACGTTACGTGTTCTAAAGATGGTG GAACCATGGCTCAGAGGAAACCAATTTTGCATAAAAATTCTAAAT CCCTATATGCCGAGTGTGGTAGAAACTTTGGAGCAAATGCAAAGA AAACATGGAGGAATGCTAGTGCGAAATCCACTCTCAAGAAACTCC ACTCATGAAATGTACTGGGTTTCATGTGGAACAGGAAACATTGTG TCAGCAGTAAACATGACATCTAGAATGCTGCTAAATCGATTCACA ATGGCTCACAGGAAGCCAACATATGAAAGAGACGTGGACTTAGGC GCTGGAACAAGACATGTGGCAGTAGAACCAGAGGTGGCCAACCTA GATATCATTGGCCAGAGGATAGAGAATATAAAAAATGAACACAA ATCAACATGGCATTATGATGAGGACAATCCATACAAAACATGGGC CTATCATGGATCATATGAGGTCAAGCCATCAGGATCAGCCTCATCC ATGGTCAATGGTGTGGTGAGACTGCTAACCAAACCATGGGATGTC ATTCCCATGGTCACACAAATAGCCATGACTGACACCACACCCTTTG GACAACAGAGGGTGTTTAAAGAGAAAGTTGACACGCGTACACCAA AAGCGAAACGAGGCACAGCACAAATTATGGAGGTGACAGCCAGG TGGTTATGGGGTTTTCTCTCTAGAAACAAAAAACCCAGAATCTGCA CAAGAGAGGAGTTCACAAGAAAAGTCAGGTCAAACGCAGCTATTG GAGCAGTGTTCGTTGATGAAAATCAATGGAACTCAGCAAAAGAGG CAGTGGAAGATGAACGGTTCTGGGACCTTGTGCACAGAGAGAGGG AGCTTCATAAACAAGGAAAATGTGCCACGTGTGTCTACAACATGA TGGGAAAGAGAGAGAAAAAATTAGGAGAGTTCGGAAAGGCAAAA GGAAGTCGCGCAATATGGTACATGTGGTTGGGAGCGCGCTTTTTA GAGTTTGAAGCCCTTGGTTTCATGAATGAAGATCACTGGTTCAGCA GAGAGAATTCACTCAGTGGAGTGGAAGGAGAAGGACTCCACAAA CTTGGATACATACTCAGAGACATATCAAAGATTCCAGGGGGAAAT ATGTATGCAGATGACACAGCCGGATGGGACACAAGAATAACAGA GGATGATCTTCAGAATGAGGCCAAAATCACTGACATCATGGAACC TGAACATGCCCTATTGGCCACGTCAATCTTTAAGCTAACCTACCAA AACAAGGTAGTAAGGGTGCAGAGACCAGCGAAAAATGGAACCGT GATGGATGTCATATCCAGACGTGACCAGAGAGGAAGTGGACAGGT TGGAACCTATGGCTTAAACACCTTCACCAACATGGAGGCCCAACT AATAAGACAAATGGAGTCTGAGGGAATCTTTTCACCCAGCGAATT GGAAACCCCAAATCTAGCCGAAAGAGTCCTCGACTGGTTGAAAAA ACATGGCACCGAGAGGCTGAAAAGAATGGCAATCAGTGGAGATG ACTGTGTGGTGAAACCAATCGATGACAGATTTGCAACAGCCTTAA CAGCTTTGAATGACATGGGAAAGGTAAGAAAAGACATACCGCAAT GGGAACCTTCAAAAGGATGGAATGATTGGCAACAAGTGCCTTTCT GTTCACACCATTTCCACCAGCTGATTATGAAGGATGGGAGGGAGA TAGTGGTGCCATGCCGCAACCAAGATGAACTTGTAGGTAGGGCCA GAGTATCACAAGGCGCCGGATGGAGCTTGAGAGAAACTGCATGCC TAGGCAAGTCATATGCACAAATGTGGCAGCTGATGTACTTCCACA GGAGAGACTTGAGATTAGCGGCTAATGCTATCTGTTCAGCCGTTCC AGTTGATTGGGTCCCAACCAGCCGCACCACCTGGTCGATCCATGCC CACCATCAATGGATGACAACAGAAGACATGTTGTCAGTGTGGAAT AGGGTTTGGATAGAGGAAAACCCATGGATGGAGGACAAGACTCAT GTGTCCAGTTGGGAAGACGTTCCATACCTAGGAAAAAGGGAAGAT CAATGGTGTGGTTCCCTAATAGGCTTAACAGCACGAGCCACCTGG GCCACCAACATACAAGTGGCCATAAACCAAGTGAGAAGGCTCATT GGGAATGAGAATTATCTAGACTTCATGACATCAATGAAGAGATTC AAAAACGAGAGTGATCCCGAAGGGGCACTCTGGTAAGCCAACTCA TTCACAAAATAAAGGAAAATAAAAAATCAAACAAGGCAAGAAGT CAGGCCGGATTAAGCCATAGCACGGTAAGAGCTATGCTGCCTGTG AGCCCCGTCCAAGGACGTAAAATGAAGTCAGGCCGAAAGCCACG GTTCGAGCAAGCCGTGCTGCCTGTAGCTCCATCGTGGGGATGTAA AAACCCGGGAGGCTGCAAACCATGGAAGCTGTACGCATGGGGTAG CAGACTAGTGGTTAGAGGAGACCCCTCCCAAGACACAACGCAGCA GCGGGGCCCAACACCAGGGGAAGCTGTACCCTGGTGGTAAGGACT AGAGGTTAGAGGAGACCCCCCGCACAACAACAAACAGCATATTGA CGCTGGGAGAGACCAGAGATCCTGCTGTCTCTACAGCATCATTCC AGGCACAGAACGCCAAAAAATGGAATGGTGCTGTTGAATCAACAG GTTCT DEN-2 AGTTGTTAGTCTACGTGGACCGACAAAGACAGATTCTTTGAGGGA 224 (NC_001474.2) GCTAAGCTCAACGTAGTTCTAACAGTTTTTTAATTAGAGAGCAGAT CTCTGATGAATAACCAACGGAAAAAGGCGAAAAACACGCCTTTCA ATATGCTGAAACGCGAGAGAAACCGCGTGTCGACTGTGCAACAGC TGACAAAGAGATTCTCACTTGGAATGCTGCAGGGACGAGGACCAT TAAAACTGTTCATGGCCCTGGTGGCGTTCCTTCGTTTCCTAACAAT CCCACCAACAGCAGGGATATTGAAGAGATGGGGAACAATTAAAA AATCAAAAGCTATTAATGTTTTGAGAGGGTTCAGGAAAGAGATTG GAAGGATGCTGAACATCTTGAATAGGAGACGCAGATCTGCAGGCA TGATCATTATGCTGATTCCAACAGTGATGGCGTTCCATTTAACCAC ACGTAACGGAGAACCACACATGATCGTCAGCAGACAAGAGAAAG GGAAAAGTCTTCTGTTTAAAACAGAGGATGGCGTGAACATGTGTA CCCTCATGGCCATGGACCTTGGTGAATTGTGTGAAGACACAATCA CGTACAAGTGTCCCCTTCTCAGGCAGAATGAGCCAGAAGACATAG ACTGTTGGTGCAACTCTACGTCCACGTGGGTAACTTATGGGACGTG TACCACCATGGGAGAACATAGAAGAGAAAAAAGATCAGTGGCAC TCGTTCCACATGTGGGAATGGGACTGGAGACACGAACTGAAACAT GGATGTCATCAGAAGGGGCCTGGAAACATGTCCAGAGAATTGAAA CTTGGATCTTGAGACATCCAGGCTTCACCATGATGGCAGCAATCCT GGCATACACCATAGGAACGACACATTTCCAAAGAGCCCTGATTTT CATCTTACTGACAGCTGTCACTCCTTCAATGACAATGCGTTGCATA GGAATGTCAAATAGAGACTTTGTGGAAGGGGTTTCAGGAGGAAGC TGGGTTGACATAGTCTTAGAACATGGAAGCTGTGTGACGACGATG GCAAAAAACAAACCAACATTGGATTTTGAACTGATAAAAACAGAA GCCAAACAGCCTGCCACCCTAAGGAAGTACTGTATAGAGGCAAAG CTAACCAACACAACAACAGAATCTCGCTGCCCAACACAAGGGGAA CCCAGCCTAAATGAAGAGCAGGACAAAAGGTTCGTCTGCAAACAC TCCATGGTAGACAGAGGATGGGGAAATGGATGTGGACTATTTGGA AAGGGAGGCATTGTGACCTGTGCTATGTTCAGATGCAAAAAGAAC ATGGAAGGAAAAGTTGTGCAACCAGAAAACTTGGAATACACCATT GTGATAACACCTCACTCAGGGGAAGAGCATGCAGTCGGAAATGAC ACAGGAAAACATGGCAAGGAAATCAAAATAACACCACAGAGTTC CATCACAGAAGCAGAATTGACAGGTTATGGCACTGTCACAATGGA GTGCTCTCCAAGAACGGGCCTCGACTTCAATGAGATGGTGTTGCTG CAGATGGAAAATAAAGCTTGGCTGGTGCACAGGCAATGGTTCCTA GACCTGCCGTTACCATGGTTGCCCGGAGCGGACACACAAGGGTCA AATTGGATACAGAAAGAGACATTGGTCACTTTCAAAAATCCCCAT GCGAAGAAACAGGATGTTGTTGTTTTAGGATCCCAAGAAGGGGCC ATGCACACAGCACTTACAGGGGCCACAGAAATCCAAATGTCATCA GGAAACTTACTCTTCACAGGACATCTCAAGTGCAGGCTGAGAATG GACAAGCTACAGCTCAAAGGAATGTCATACTCTATGTGCACAGGA AAGTTTAAAGTTGTGAAGGAAATAGCAGAAACACAACATGGAAC AATAGTTATCAGAGTGCAATATGAAGGGGACGGCTCTCCATGCAA GATCCCTTTTGAGATAATGGATTTGGAAAAAAGACATGTCTTAGGT CGCCTGATTACAGTCAACCCAATTGTGACAGAAAAAGATAGCCCA GTCAACATAGAAGCAGAACCTCCATTCGGAGACAGCTACATCATC ATAGGAGTAGAGCCGGGACAACTGAAGCTCAACTGGTTTAAGAAA GGAAGTTCTATCGGCCAAATGTTTGAGACAACAATGAGGGGGGCG AAGAGAATGGCCATTTTAGGTGACACAGCCTGGGATTTTGGATCC TTGGGAGGAGTGTTTACATCTATAGGAAAGGCTCTCCACCAAGTCT TTGGAGCAATCTATGGAGCTGCCTTCAGTGGGGTTTCATGGACTAT GAAAATCCTCATAGGAGTCATTATCACATGGATAGGAATGAATTC ACGCAGCACCTCACTGTCTGTGACACTAGTATTGGTGGGAATTGTG ACACTGTATTTGGGAGTCATGGTGCAGGCCGATAGTGGTTGCGTTG TGAGCTGGAAAAACAAAGAACTGAAATGTGGCAGTGGGATTTTCA TCACAGACAACGTGCACACATGGACAGAACAATACAAGTTCCAAC CAGAATCCCCTTCAAAACTAGCTTCAGCTATCCAGAAAGCCCATG AAGAGGGCATTTGTGGAATCCGCTCAGTAACAAGACTGGAGAATC TGATGTGGAAACAAATAACACCAGAATTGAATCACATTCTATCAG AAAATGAGGTGAAGTTAACTATTATGACAGGAGACATCAAAGGAA TCATGCAGGCAGGAAAACGATCTCTGCGGCCTCAGCCCACTGAGC TGAAGTATTCATGGAAAACATGGGGCAAAGCAAAAATGCTCTCTA CAGAGTCTCATAACCAGACCTTTCTCATTGATGGCCCCGAAACAGC AGAATGCCCCAACACAAATAGAGCTTGGAATTCGTTGGAAGTTGA AGACTATGGCTTTGGAGTATTCACCACCAATATATGGCTAAAATTG AAAGAAAAACAGGATGTATTCTGCGACTCAAAACTCATGTCAGCG GCCATAAAAGACAACAGAGCCGTCCATGCCGATATGGGTTATTGG ATAGAAAGTGCACTCAATGACACATGGAAGATAGAGAAAGCCTCT TTCATTGAAGTTAAAAACTGCCACTGGCCAAAATCACACACCCTCT GGAGCAATGGAGTGCTAGAAAGTGAGATGATAATTCCAAAGAATC TCGCTGGACCAGTGTCTCAACACAACTATAGACCAGGCTACCATA CACAAATAACAGGACCATGGCATCTAGGTAAGCTTGAGATGGACT TTGATTTCTGTGATGGAACAACAGTGGTAGTGACTGAGGACTGCG GAAATAGAGGACCCTCTTTGAGAACAACCACTGCCTCTGGAAAAC TCATAACAGAATGGTGCTGCCGATCTTGCACATTACCACCGCTAAG ATACAGAGGTGAGGATGGGTGCTGGTACGGGATGGAAATCAGACC ATTGAAGGAGAAAGAAGAGAATTTGGTCAACTCCTTGGTCACAGC TGGACATGGGCAGGTCGACAACTTTTCACTAGGAGTCTTGGGAAT GGCATTGTTCCTGGAGGAAATGCTTAGGACCCGAGTAGGAACGAA ACATGCAATACTACTAGTTGCAGTTTCTTTTGTGACATTGATCACA GGGAACATGTCCTTTAGAGACCTGGGAAGAGTGATGGTTATGGTA GGCGCCACTATGACGGATGACATAGGTATGGGCGTGACTTATCTT GCCCTACTAGCAGCCTTCAAAGTCAGACCAACTTTTGCAGCTGGAC TACTCTTGAGAAAGCTGACCTCCAAGGAATTGATGATGACTACTAT AGGAATTGTACTCCTCTCCCAGAGCACCATACCAGAGACCATTCTT GAGTTGACTGATGCGTTAGCCTTAGGCATGATGGTCCTCAAAATG GTGAGAAATATGGAAAAGTATCAATTGGCAGTGACTATCATGGCT ATCTTGTGCGTCCCAAACGCAGTGATATTACAAAACGCATGGAAA GTGAGTTGCACAATATTGGCAGTGGTGTCCGTTTCCCCACTGCTCT TAACATCCTCACAGCAAAAAACAGATTGGATACCATTAGCATTGA CGATCAAAGGTCTCAATCCAACAGCTATTTTTCTAACAACCCTCTC AAGAACCAGCAAGAAAAGGAGCTGGCCATTAAATGAGGCTATCAT GGCAGTCGGGATGGTGAGCATTTTAGCCAGTTCTCTCCTAAAAAAT GATATTCCCATGACAGGACCATTAGTGGCTGGAGGGCTCCTCACT GTGTGCTACGTGCTCACTGGACGATCGGCCGATTTGGAACTGGAG AGAGCAGCCGATGTCAAATGGGAAGACCAGGCAGAGATATCAGG AAGCAGTCCAATCCTGTCAATAACAATATCAGAAGATGGTAGCAT GTCGATAAAAAATGAAGAGGAAGAACAAACACTGACCATACTCAT TAGAACAGGATTGCTGGTGATCTCAGGACTTTTTCCTGTATCAATA CCAATCACGGCAGCAGCATGGTACCTGTGGGAAGTGAAGAAACAA CGGGCCGGAGTATTGTGGGATGTTCCTTCACCCCCACCCATGGGA AAGGCTGAACTGGAAGATGGAGCCTATAGAATTAAGCAAAAAGG GATTCTTGGATATTCCCAGATCGGAGCCGGAGTTTACAAAGAAGG AACATTCCATACAATGTGGCATGTCACACGTGGCGCTGTTCTAATG CATAAAGGAAAGAGGATTGAACCATCATGGGCGGACGTCAAGAA AGACCTAATATCATATGGAGGAGGCTGGAAGTTAGAAGGAGAATG GAAGGAAGGAGAAGAAGTCCAGGTATTGGCACTGGAGCCTGGAA AAAATCCAAGAGCCGTCCAAACGAAACCTGGTCTTTTCAAAACCA ACGCCGGAACAATAGGTGCTGTATCTCTGGACTTTTCTCCTGGAAC GTCAGGATCTCCAATTATCGACAAAAAAGGAAAAGTTGTGGGTCT TTATGGTAATGGTGTTGTTACAAGGAGTGGAGCATATGTGAGTGCT ATAGCCCAGACTGAAAAAAGCATTGAAGACAACCCAGAGATCGA AGATGACATTTTCCGAAAGAGAAGACTGACCATCATGGACCTCCA CCCAGGAGCGGGAAAGACGAAGAGATACCTTCCGGCCATAGTCAG AGAAGCTATAAAACGGGGTTTGAGAACATTAATCTTGGCCCCCAC TAGAGTTGTGGCAGCTGAAATGGAGGAAGCCCTTAGAGGACTTCC AATAAGATACCAGACCCCAGCCATCAGAGCTGAGCACACCGGGCG GGAGATTGTGGACCTAATGTGTCATGCCACATTTACCATGAGGCTG CTATCACCAGTTAGAGTGCCAAACTACAACCTGATTATCATGGAC GAAGCCCATTTCACAGACCCAGCAAGTATAGCAGCTAGAGGATAC ATCTCAACTCGAGTGGAGATGGGTGAGGCAGCTGGGATTTTTATG ACAGCCACTCCCCCGGGAAGCAGAGACCCATTTCCTCAGAGCAAT GCACCAATCATAGATGAAGAAAGAGAAATCCCTGAACGTTCGTGG AATTCCGGACATGAATGGGTCACGGATTTTAAAGGGAAGACTGTT TGGTTCGTTCCAAGTATAAAAGCAGGAAATGATATAGCAGCTTGC CTGAGGAAAAATGGAAAGAAAGTGATACAACTCAGTAGGAAGAC CTTTGATTCTGAGTATGTCAAGACTAGAACCAATGATTGGGACTTC GTGGTTACAACTGACATTTCAGAAATGGGTGCCAATTTCAAGGCT GAGAGGGTTATAGACCCCAGACGCTGCATGAAACCAGTCATACTA ACAGATGGTGAAGAGCGGGTGATTCTGGCAGGACCTATGCCAGTG ACCCACTCTAGTGCAGCACAAAGAAGAGGGAGAATAGGAAGAAA TCCAAAAAATGAGAATGACCAGTACATATACATGGGGGAACCTCT GGAAAATGATGAAGACTGTGCACACTGGAAAGAAGCTAAAATGCT CCTAGATAACATCAACACGCCAGAAGGAATCATTCCTAGCATGTT CGAACCAGAGCGTGAAAAGGTGGATGCCATTGATGGCGAATACCG CTTGAGAGGAGAAGCAAGGAAAACCTTTGTAGACTTAATGAGAAG AGGAGACCTACCAGTCTGGTTGGCCTACAGAGTGGCAGCTGAAGG CATCAACTACGCAGACAGAAGGTGGTGTTTTGATGGAGTCAAGAA CAACCAAATCCTAGAAGAAAACGTGGAAGTTGAAATCTGGACAAA AGAAGGGGAAAGGAAGAAATTGAAACCCAGATGGTTGGATGCTA GGATCTATTCTGACCCACTGGCGCTAAAAGAATTTAAGGAATTTGC AGCCGGAAGAAAGTCTCTGACCCTGAACCTAATCACAGAAATGGG TAGGCTCCCAACCTTCATGACTCAGAAGGCAAGAGACGCACTGGA CAACTTAGCAGTGCTGCACACGGCTGAGGCAGGTGGAAGGGCGTA CAACCATGCTCTCAGTGAACTGCCGGAGACCCTGGAGACATTGCT TTTACTGACACTTCTGGCTACAGTCACGGGAGGGATCTTTTTATTC TTGATGAGCGGAAGGGGCATAGGGAAGATGACCCTGGGAATGTGC TGCATAATCACGGCTAGCATCCTCCTATGGTACGCACAAATACAG CCACACTGGATAGCAGCTTCAATAATACTGGAGTTTTTTCTCATAG TTTTGCTTATTCCAGAACCTGAAAAACAGAGAACACCCCAAGACA ACCAACTGACCTACGTTGTCATAGCCATCCTCACAGTGGTGGCCGC AACCATGGCAAACGAGATGGGTTTCCTAGAAAAAACGAAGAAAG ATCTCGGATTGGGAAGCATTGCAACCCAGCAACCCGAGAGCAACA TCCTGGACATAGATCTACGTCCTGCATCAGCATGGACGCTGTATGC CGTGGCCACAACATTTGTTACACCAATGTTGAGACATAGCATTGA AAATTCCTCAGTGAATGTGTCCCTAACAGCTATAGCCAACCAAGC CACAGTGTTAATGGGTCTCGGGAAAGGATGGCCATTGTCAAAGAT GGACATCGGAGTTCCCCTTCTCGCCATTGGATGCTACTCACAAGTC AACCCCATAACTCTCACAGCAGCTCTTTTCTTATTGGTAGCACATT ATGCCATCATAGGGCCAGGACTCCAAGCAAAAGCAACCAGAGAA GCTCAGAAAAGAGCAGCGGCGGGCATCATGAAAAACCCAACTGTC GATGGAATAACAGTGATTGACCTAGATCCAATACCTTATGATCCA AAGTTTGAAAAGCAGTTGGGACAAGTAATGCTCCTAGTCCTCTGC GTGACTCAAGTATTGATGATGAGGACTACATGGGCTCTGTGTGAG GCTTTAACCTTAGCTACCGGGCCCATCTCCACATTGTGGGAAGGAA ATCCAGGGAGGTTTTGGAACACTACCATTGCGGTGTCAATGGCTA ACATTTTTAGAGGGAGTTACTTGGCCGGAGCTGGACTTCTCTTTTC TATTATGAAGAACACAACCAACACAAGAAGGGGAACTGGCAACA TAGGAGAGACGCTTGGAGAGAAATGGAAAAGCCGATTGAACGCA TTGGGAAAAAGTGAATTCCAGATCTACAAGAAAAGTGGAATCCAG GAAGTGGATAGAACCTTAGCAAAAGAAGGCATTAAAAGAGGAGA AACGGACCATCACGCTGTGTCGCGAGGCTCAGCAAAACTGAGATG GTTCGTTGAGAGAAACATGGTCACACCAGAAGGGAAAGTAGTGGA CCTCGGTTGTGGCAGAGGAGGCTGGTCATACTATTGTGGAGGACT AAAGAATGTAAGAGAAGTCAAAGGCCTAACAAAAGGAGGACCAG GACACGAAGAACCCATCCCCATGTCAACATATGGGTGGAATCTAG TGCGTCTTCAAAGTGGAGTTGACGTTTTCTTCATCCCGCCAGAAAA GTGTGACACATTATTGTGTGACATAGGGGAGTCATCACCAAATCC CACAGTGGAAGCAGGACGAACACTCAGAGTCCTTAACTTAGTAGA AAATTGGTTGAACAACAACACTCAATTTTGCATAAAGGTTCTCAAC CCATATATGCCCTCAGTCATAGAAAAAATGGAAGCACTACAAAGG AAATATGGAGGAGCCTTAGTGAGGAATCCACTCTCACGAAACTCC ACACATGAGATGTACTGGGTATCCAATGCTTCCGGGAACATAGTG TCATCAGTGAACATGATTTCAAGGATGTTGATCAACAGATTTACAA TGAGATACAAGAAAGCCACTTACGAGCCGGATGTTGACCTCGGAA GCGGAACCCGTAACATCGGGATTGAAAGTGAGATACCAAACCTAG ATATAATTGGGAAAAGAATAGAAAAAATAAAGCAAGAGCATGAA ACATCATGGCACTATGACCAAGACCACCCATACAAAACGTGGGCA TACCATGGTAGCTATGAAACAAAACAGACTGGATCAGCATCATCC ATGGTCAACGGAGTGGTCAGGCTGCTGACAAAACCTTGGGACGTC GTCCCCATGGTGACACAGATGGCAATGACAGACACGACTCCATTT GGACAACAGCGCGTTTTTAAAGAGAAAGTGGACACGAGAACCCA AGAACCGAAAGAAGGCACGAAGAAACTAATGAAAATAACAGCAG AGTGGCTTTGGAAAGAATTAGGGAAGAAAAAGACACCCAGGATG TGCACCAGAGAAGAATTCACAAGAAAGGTGAGAAGCAATGCAGC CTTGGGGGCCATATTCACTGATGAGAACAAGTGGAAGTCGGCACG TGAGGCTGTTGAAGATAGTAGGTTTTGGGAGCTGGTTGACAAGGA AAGGAATCTCCATCTTGAAGGAAAGTGTGAAACATGTGTGTACAA CATGATGGGAAAAAGAGAGAAGAAGCTAGGGGAATTCGGCAAGG CAAAAGGCAGCAGAGCCATATGGTACATGTGGCTTGGAGCACGCT TCTTAGAGTTTGAAGCCCTAGGATTCTTAAATGAAGATCACTGGTT CTCCAGAGAGAACTCCCTGAGTGGAGTGGAAGGAGAAGGGCTGC ACAAGCTAGGTTACATTCTAAGAGACGTGAGCAAGAAAGAGGGA GGAGCAATGTATGCCGATGACACCGCAGGATGGGATACAAGAATC ACACTAGAAGACCTAAAAAATGAAGAAATGGTAACAAACCACAT GGAAGGAGAACACAAGAAACTAGCCGAGGCCATTTTCAAACTAAC GTACCAAAACAAGGTGGTGCGTGTGCAAAGACCAACACCAAGAG GCACAGTAATGGACATCATATCGAGAAGAGACCAAAGAGGTAGT GGACAAGTTGGCACCTATGGACTCAATACTTTCACCAATATGGAA GCCCAACTAATCAGACAGATGGAGGGAGAAGGAGTCTTTAAAAGC ATTCAGCACCTAACAATCACAGAAGAAATCGCTGTGCAAAACTGG TTAGCAAGAGTGGGGCGCGAAAGGTTATCAAGAATGGCCATCAGT GGAGATGATTGTGTTGTGAAACCTTTAGATGACAGGTTCGCAAGC GCTTTAACAGCTCTAAATGACATGGGAAAGATTAGGAAAGACATA CAACAATGGGAACCTTCAAGAGGATGGAATGATTGGACACAAGTG CCCTTCTGTTCACACCATTTCCATGAGTTAATCATGAAAGACGGTC GCGTACTCGTTGTTCCATGTAGAAACCAAGATGAACTGATTGGCA GAGCCCGAATCTCCCAAGGAGCAGGGTGGTCTTTGCGGGAGACGG CCTGTTTGGGGAAGTCTTACGCCCAAATGTGGAGCTTGATGTACTT CCACAGACGCGACCTCAGGCTGGCGGCAAATGCTATTTGCTCGGC AGTACCATCACATTGGGTTCCAACAAGTCGAACAACCTGGTCCAT ACATGCTAAACATGAATGGATGACAACGGAAGACATGCTGACAGT CTGGAACAGGGTGTGGATTCAAGAAAACCCATGGATGGAAGACA AAACTCCAGTGGAATCATGGGAGGAAATCCCATACTTGGGGAAAA GAGAAGACCAATGGTGCGGCTCATTGATTGGGTTAACAAGCAGGG CCACCTGGGCAAAGAACATCCAAGCAGCAATAAATCAAGTTAGAT CCCTTATAGGCAATGAAGAATACACAGATTACATGCCATCCATGA AAAGATTCAGAAGAGAAGAGGAAGAAGCAGGAGTTCTGTGGTAG AAAGCAAAACTAACATGAAACAAGGCTAGAAGTCAGGTCGGATT AAGCCATAGTACGGAAAAAACTATGCTACCTGTGAGCCCCGTCCA AGGACGTTAAAAGAAGTCAGGCCATCATAAATGCCATAGCTTGAG TAAACTATGCAGCCTGTAGCTCCACCTGAGAAGGTGTAAAAAATC CGGGAGGCCACAAACCATGGAAGCTGTACGCATGGCGTAGTGGAC TAGCGGTTAGAGGAGACCCCTCCCTTACAAATCGCAGCAACAATG GGGGCCCAAGGCGAGATGAAGCTGTAGTCTCGCTGGAAGGACTAG AGGTTAGAGGAGACCCCCCCGAAACAAAAAACAGCATATTGACGC TGGGAAAGACCAGAGATCCTGCTGTCTCCTCAGCATCATTCCAGG CACAGAACGCCAGAAAATGGAATGGTGCTGTTGAATCAACAGGTT CT DEN-3 AGTTGTTAGTCTACGTGGACCGACAAGAACAGTTTCGACTCGGAA 225 (NC_001475.2) GCTTGCTTAACGTAGTGCTGACAGTTTTTTATTAGAGAGCAGATCT CTGATGAACAACCAACGGAAGAAGACGGGAAAACCGTCTATCAAT ATGCTGAAACGCGTGAGAAACCGTGTGTCAACTGGATCACAGTTG GCGAAGAGATTCTCAAAAGGACTGCTGAACGGCCAGGGACCAATG AAATTGGTTATGGCGTTCATAGCTTTCCTCAGATTTCTAGCCATTC CACCAACAGCAGGAGTCTTGGCTAGATGGGGAACCTTCAAGAAGT CGGGGGCCATTAAGGTCCTGAAAGGCTTCAAGAAGGAGATCTCAA ACATGCTGAGCATAATCAACCAACGGAAAAAGACATCGCTCTGTC TCATGATGATATTGCCAGCAGCACTTGCTTTCCACTTGACTTCACG AGATGGAGAGCCGCGCATGATTGTGGGGAAGAATGAAAGAGGTA AATCCCTACTTTTTAAGACAGCCTCTGGAATCAACATGTGCACACT CATAGCCATGGATTTGGGAGAGATGTGTGATGACACGGTCACTTA CAAATGCCCCCACATTACCGAAGTGGAACCTGAAGACATTGACTG CTGGTGCAACCTTACATCAACATGGGTGACTTATGGAACGTGCAA TCAAGCTGGAGAGCATAGACGCGACAAGAGATCAGTGGCGTTAGC TCCCCATGTCGGCATGGGACTGGACACACGCACCCAAACCTGGAT GTCGGCTGAAGGAGCTTGGAGACAAGTCGAGAAGGTAGAGACAT GGGCCCTTAGGCACCCAGGGTTCACCATACTAGCCCTATTTCTCGC CCATTACATAGGCACTTCCCTGACCCAGAAGGTGGTTATTTTCATA TTATTAATGCTGGTCACCCCATCCATGACAATGAGATGTGTGGGAG TAGGAAACAGAGATTTTGTGGAAGGGCTATCAGGAGCTACGTGGG TTGACGTGGTGCTCGAGCACGGGGGGTGTGTGACTACCATGGCTA AGAACAAGCCCACGCTGGATATAGAGCTTCAGAAGACCGAGGCCA CCCAACTGGCGACCCTAAGGAAGCTATGCATTGAGGGGAAAATTA CCAACATAACAACTGACTCAAGATGTCCTACCCAAGGGGAAGCGG TTTTGCCTGAGGAGCAGGACCAGAACTACGTGTGTAAGCATACAT ACGTAGACAGAGGTTGGGGGAACGGTTGTGGTTTGTTTGGCAAAG GAAGCTTGGTAACATGTGCGAAATTTCAATGCCTGGAACCAATAG AGGGAAAAGTGGTGCAATATGAGAACCTCAAATACACCGTCATCA TTACAGTGCACACAGGAGACCAACACCAGGTGGGAAATGAAACG CAAGGAGTCACGGCTGAGATAACACCTCAGGCATCAACCACTGAA GCCATCTTGCCTGAATATGGAACCCTTGGGCTAGAATGCTCACCAC GGACAGGTTTGGATTTCAATGAAATGATCTTACTAACAATGAAGA ACAAAGCATGGATGGTACATAGACAATGGTTCTTTGACCTACCTCT ACCATGGGCATCAGGAGCTACAACAGAAACACCAACCTGGAACA GGAAGGAGCTTCTTGTGACATTCAAAAACGCACATGCGAAAAAAC AAGAAGTAGTTGTCCTTGGATCGCAAGAGGGAGCAATGCATACCG CACTGACAGGAGCTACAGAAATCCAAAACTCAGGAGGCACAAGC ATTTTCGCGGGGCACTTAAAATGTAGACTTAAGATGGACAAATTG GAACTCAAGGGGATGAGCTATGCAATGTGCACGAATACCTTTGTG TTGAAGAAAGAAGTCTCAGAAACGCAGCACGGGACAATACTCATT AAGGTTGAGTACAAAGGGGAAGATGCACCTTGCAAGATTCCCTTT TCCACAGAGGATGGACAAGGGAAAGCTCATAATGGCAGACTGATC ACAGCCAACCCTGTGGTGACTAAGAAGGAGGAGCCTGTCAATATT GAGGCTGAACCTCCTTTTGGGGAAAGCAATATAGTAATTGGAATT GGAGACAACGCCTTGAAAATCAACTGGTACAAGAAGGGGAGCTC GATTGGGAAGATGTTCGAGGCCACTGAAAGGGGTGCAAGGCGCAT GGCCATCTTGGGAGACACAGCTTGGGACTTTGGATCAGTGGGTGG TGTTCTGAACTCATTAGGCAAAATGGTGCACCAAATATTTGGAAGT GCTTATACAGCCCTGTTCAGTGGAGTCTCTTGGGTGATGAAAATTG GAATAGGTGTCCTCTTGACTTGGATAGGGTTGAATTCAAAAAACA CATCCATGTCATTTTCATGCATTGCGATAGGAATCATTACACTCTA TCTGGGAGCTGTGGTACAAGCTGACATGGGGTGTGTCATAAACTG GAAGGGCAAAGAACTCAAATGTGGAAGCGGAATTTTCGTCACCAA TGAGGTCCATACCTGGACAGAGCAATACAAATTCCAAGCAGACTC CCCAAAAAGATTGGCAACAGCCATTGCAGGCGCCTGGGAGAATGG AGTGTGTGGAATTAGGTCAACAACCAGAATGGAGAATCTCTTGTG GAAGCAAATAGCCAATGAACTGAACTACATATTATGGGAAAACAA TATCAAATTAACGGTAGTTGTGGGCGATACACTTGGGGTCTTAGA GCAAGGGAAAAGAACACTAACACCACAACCCATGGAGCTAAAAT ACTCATGGAAAACGTGGGGAAAGGCAAAAATAGTGACAGCTGAA ACACAAAATTCCTCTTTCATAATAGACGGGCCAAACACACCGGAG TGTCCAAGTGCCTCAAGAGCATGGAATGTGTGGGAGGTGGAAGAT TACGGGTTCGGAGTCTTCACAACCAACATATGGCTGAAACTCCGA GAGGTCTACACCCAACTATGTGACCATAGGCTAATGTCGGCAGCT GTCAAGGATGAGAGGGCCGTGCATGCCGACATGGGCTACTGGATA GAAAGCCAAAAGAATGGAAGTTGGAAGCTAGAAAAAGCATCCCT CATAGAGGTAAAAACCTGCACATGGCCAAAATCACACACTCTCTG GACTAATGGTGTGCTAGAGAGTGACATGATCATCCCAAAGAGTCT AGCTGGTCCTATCTCACAACACAACTACAGGCCCGGGTACCACAC CCAAACGGCAGGACCCTGGCACTTAGGAAAATTGGAGCTGGACTT CAACTACTGTGAAGGAACAACAGTTGTCATCACAGAAAGCTGTGG GACAAGAGGCCCATCATTGAGAACAACAACAGTGTCAGGGAAGTT GATACACGAATGGTGTTGCCGCTCGTGCACACTTCCCCCCCTGCGA TACATGGGAGAAGACGGCTGCTGGTATGGCATGGAAATCAGACCC ATCAGTGAGAAAGAAGAGAACATGGTAAAGTCTTTAGTCTCAGCG GGAAGTGGAAAGGTGGACAACTTCACAATGGGTGTCTTGTGTTTG GCAATCCTCTTTGAAGAGGTGTTGAGAGGAAAATTTGGGAAGAAA CACATGATTGCAGGGGTTTTCTTTACGTTTGTGCTCCTTCTCTCAGG GCAAATAACATGGAGAGACATGGCGCACACACTAATAATGATCGG GTCCAACGCCTCTGACAGGATGGGAATGGGCGTCACCTACCTAGC TCTAATTGCAACATTTAAAATCCAGCCATTCTTGGCTTTGGGATTT TTCCTAAGAAAGCTGACATCTAGAGAAAATTTATTGTTAGGAGTTG GGTTGGCCATGGCAACAACGTTACAACTGCCAGAGGACATTGAAC AAATGGCAAATGGAGTCGCTCTGGGGCTCATGGCTCTTAAACTGA TAACACAATTTGAAACATACCAATTGTGGACGGCATTAGTCTCCTT AACGTGTTCAAACACAATTTTTACGTTGACTGTTGCCTGGAGAACA GCCACTCTGATTTTGGCCGGAGTTTCGCTTTTACCAGTGTGCCAGT CTTCAAGCATGAGGAAAACAGATTGGCTCCCAATGACAGTGGCAG CTATGGGAGTTCCACCCCTTCCACTTTTTATTTTTAGCTTGAAAGAC ACACTCAAAAGGAGAAGCTGGCCACTGAATGAAGGGGTGATGGCT GTTGGGCTTGTGAGCATTCTGGCCAGTTCTCTCCTTAGAAATGATG TGCCCATGGCTGGACCATTAGTGGCCGGGGGCTTGCTGATAGCGT GCTACGTCATAACTGGCACGTCAGCGGACCTCACTGTAGAAAAAG CCCCAGATGTAACATGGGAGGAAGAGGCTGAGCAGACAGGAGTG TCCCACAACTTAATGATCACAGTTGATGATGATGGAACAATGAGA ATAAAAGATGATGAGACTGAGAACATCCTAACAGTGCTTTTAAAA ACAGCATTACTAATAGTATCAGGCATTTTTCCATACTCCATACCCG CAACATTGTTGGTCTGGCACACTTGGCAAAAACAAACCCAAAGAT CCGGCGTTTTATGGGACGTACCCAGCCCCCCAGAGACACAGAAAG CAGAACTGGAAGAAGGGGTTTATAGGATCAAACAGCAAGGAATTT TTGGGAAAACCCAAGTAGGGGTTGGAGTACAGAAAGAAGGAGTC TTCCACACCATGTGGCACGTCACAAGAGGGGCAGTGTTGACACAT AATGGGAAAAGACTGGAACCAAACTGGGCTAGTGTGAAAAAAGA TCTGATTTCATATGGAGGAGGATGGAGACTGAGCGCACAATGGCA AAAGGGGGAGGAGGTGCAGGTTATTGCCGTAGAGCCAGGGAAGA ACCCAAAGAACTTTCAAACCACGCCAGGCACTTTCCAGACTACTA CAGGGGAAATAGGAGCAATTGCACTGGATTTCAAGCCTGGAACTT CAGGATCTCCTATCATAAATAGAGAGGGAAAGGTAGTGGGACTGT ATGGCAATGGAGTGGTTACAAAGAATGGTGGCTATGTCAGCGGAA TAGCGCAAACAAATGCAGAACCAGATGGACCGACACCAGAGTTG GAAGAAGAGATGTTCAAAAAGCGAAACCTGACCATAATGGATCTT CATCCTGGGTCAGGAAAGACACGGAAATACCTTCCAGCTATTGTC AGAGAGGCAATCAAGAGACGTTTAAGAACCTTAATTTTGGCACCG ACAAGGGTGGTTGCAGCTGAGATGGAAGAAGCATTGAAAGGGCTC CCAATAAGGTACCAAACAACAGCAACAAAATCTGAACACACAGG AAGAGAGATTGTTGATCTAATGTGCCACGCAACGTTCACAATGCG TTTGCTGTCACCAGTTAGGGTTCCAAATTACAACTTGATAATAATG GATGAGGCCCATTTCACAGACCCAGCCAGTATAGCGGCTAGAGGG TACATATCAACTCGTGTTGGAATGGGAGAGGCAGCCGCAATCTTC ATGACAGCAACACCCCCTGGAACAGCTGATGCCTTTCCTCAGAGC AACGCTCCAATTCAAGATGAAGAAAGGGACATACCAGAACGCTCA TGGAATTCAGGCAATGAATGGATTACCGACTTCGCTGGGAAAACG GTGTGGTTTGTCCCTAGCATTAAAGCCGGAAATGACATAGCAAAC TGCTTGCGAAAAAACGGGAAAAAAGTCATTCAACTTAGTAGGAAG ACTTTTGACACAGAATATCAGAAGACTAAACTGAATGATTGGGAC TTTGTGGTGACAACTGACATTTCAGAAATGGGGGCCAATTTCAAA GCAGATAGAGTGATCGACCCAAGAAGATGTCTCAAACCAGTGATC TTGACAGATGGACCAGAGCGGGTGATCCTGGCCGGACCAATGCCA GTCACCGCGGCGAGTGCTGCGCAAAGGAGAGGGAGAGTTGGCAG GAACCCACAAAAAGAGAATGACCAGTACATATTCACGGGCCAGCC TCTCAACAATGATGAAGACCATGCTCACTGGACAGAAGCAAAAAT GCTGCTGGACAACATCAACACACCAGAAGGGATTATACCAGCTCT CTTTGAACCAGAAAGGGAGAAGTCAGCCGCCATAGACGGTGAGTA TCGCCTGAAGGGTGAGTCCAGGAAGACTTTCGTGGAACTCATGAG GAGGGGTGACCTTCCAGTTTGGTTAGCCCATAAAGTAGCATCAGA AGGAATCAAATACACAGATAGAAAATGGTGCTTTGATGGGCAACG CAATAATCAAATTTTAGAGGAGAACATGGATGTGGAAATTTGGAC AAAGGAAGGAGAAAAGAAAAAATTGAGACCTAGGTGGCTTGATG CCCGCACTTATTCAGATCCATTGGCACTCAAGGAATTCAAGGACTT TGCGGCTGGCAGAAAGTCAATCGCCCTTGATCTTGTGACAGAAAT AGGAAGAGTGCCTTCACATCTAGCCCACAGAACAAGAAACGCTCT GGACAATCTGGTGATGCTGCATACGTCAGAAGATGGCGGTAGGGC TTACAGGCATGCGGTGGAGGAACTACCAGAAACAATGGAAACACT CCTACTCTTGGGACTAATGATCTTGTTGACAGGTGGAGCAATGCTT TTCTTGATATCAGGTAAAGGGATTGGAAAGACTTCAATAGGACTC ATTTGTGTAATCGCTTCCAGCGGCATGTTGTGGATGGCCGAAGTTC CACTCCAATGGATCGCGTCGGCTATAGTCCTGGAGTTTTTTATGAT GGTGTTGCTCATACCAGAACCAGAAAAGCAGAGAACCCCCCAAGA CAACCAACTCGCATATGTCGTGATAGGCATACTTACATTGGCTGCA ACAATAGCAGCCAATGAAATGGGACTGCTGGAAACCACAAAGAG AGACTTAGGAATGTCTAAGGAGCCAGGTGTTGTTTCTCCAACCAG CTATTTGGATGTGGACTTGCACCCAGCATCAGCCTGGACATTGTAC GCCGTGGCCACTACAGTAATAACACCAATGTTAAGACATACCATA GAGAATTCTACAGCAAATGTGTCCCTGGCAGCTATAGCCAACCAG GCAGTGGTCCTGATGGGTTTGGACAAAGGATGGCCAATATCAAAA ATGGACTTAGGCGTGCCACTACTGGCACTGGGTTGCTATTCACAAG TGAACCCACTGACTCTAACTGCGGCAGTACTTTTGCTAATCACACA TTATGCTATCATAGGTCCAGGATTGCAAGCAAAAGCCACCCGTGA AGCTCAGAAAAGGACAGCTGCTGGAATAATGAAGAATCCAACAGT GGATGGGATAATGACAATAGACCTAGATTCTGTAATATTTGATTCA AAATTTGAAAAACAACTGGGACAGGTTATGCTCCTGGTTTTGTGCG CAGTCCAACTCTTGCTAATGAGAACATCATGGGCCTTGTGTGAAGC TTTAACTCTAGCTACAGGACCAATAACAACACTCTGGGAAGGATC ACCTGGTAAGTTCTGGAACACCACGATAGCTGTTTCCATGGCGAA CATTTTTAGAGGGAGCTATTTAGCAGGAGCTGGGCTTGCTTTTTCT ATTATGAAATCAGTTGGAACAGGAAAAAGAGGAACAGGCTCACA AGGTGAAACTTTAGGAGAAAAATGGAAAAAGAAATTAAATCAATT ATCCCGGAAAGAGTTTGACCTTTACAAGAAATCTGGAATCACTGA AGTGGATAGAACAGAAGCCAAAGAAGGGTTGAAAAGAGGAGAGA CAACACATCATGCCGTGTCCCGAGGTAGCGCAAAACTTCAATGGT TTGTGGAAAGAAACATGGTCGTTCCCGAAGGAAGAGTCATAGACT TGGGCTGTGGAAGAGGAGGCTGGTCATATTACTGTGCAGGACTGA AAAAAGTCACAGAAGTGCGAGGATACACAAAAGGCGGTCCAGGA CACGAAGAACCAGTACCTATGTCTACATATGGATGGAACATAGTT AAGTTAATGAGCGGAAAGGATGTGTTCTATCTCCCACCTGAAAAG TGTGATACCCTGTTGTGTGACATTGGAGAATCTTCACCAAGCCCAA CAGTGGAAGAGAGCAGAACTATAAGAGTTTTGAAGATGGTTGAAC CATGGCTAAAAAACAACCAGTTTTGCATTAAAGTTTTGAACCCTTA CATGCCAACTGTGATTGAGCACCTAGAAAGACTACAAAGGAAACA TGGAGGAATGCTTGTGAGAAATCCACTTTCACGAAACTCCACGCA CGAAATGTACTGGATATCTAATGGCACAGGTAACATTGTCTCTTCA GTCAACATGGTGTCTAGATTGCTACTGAACAGGTTCACGATGACA CACAGGAGACCCACCATAGAGAAAGATGTGGATTTAGGAGCAGG AACTCGACATGTTAATGCGGAACCAGAAACACCCAACATGGATGT CATTGGGGAAAGAATAAAAAGGATCAAGGAGGAGCATAATTCAA CATGGCACTATGATGACGAAAACCCCTACAAAACGTGGGCTTACC ATGGATCCTATGAAGTCAAAGCCACAGGCTCAGCCTCCTCCATGA TAAATGGAGTCGTGAAACTCCTCACCAAACCATGGGATGTGGTGC CCATGGTGACACAGATGGCAATGACAGACACAACTCCATTTGGCC AGCAGAGAGTCTTTAAAGAGAAAGTGGACACCAGGACGCCCAGG CCCATGCCAGGGACAAGAAAGGCTATGGAGATCACAGCGGAGTG GCTCTGGAGAACCCTGGGAAGGAACAAAAGACCCAGATTATGCAC AAGGGAAGAGTTTACAAAAAAGGTCAGAACTAACGCAGCCATGG GCGCCGTTTTCACAGAGGAGAACCAATGGGACAGTGCGAAAGCTG CTGTTGAGGATGAAGAATTTTGGAAACTTGTGGACAGAGAACGTG AACTCCACAAATTGGGCAAATGTGGAAGCTGCGTTTATAACATGA TGGGCAAGAGAGAGAAAAAACTTGGAGAGTTTGGCAAAGCAAAA GGCAGTAGAGCTATATGGTACATGTGGTTGGGAGCCAGGTACCTT GAGTTCGAAGCCCTTGGATTCTTAAATGAAGACCACTGGTTCTCGC GTGAAAACTCTTACAGTGGAGTAGAAGGAGAAGGACTGCACAAG CTAGGCTACATATTAAGGGACATTTCCAAGATACCCGGAGGAGCC ATGTATGCTGATGACACAGCTGGTTGGGACACAAGAATAACAGAA GATGACCTGCACAATGAGGAAAAGATCATACAGCAAATGGACCCT GAACACAGGCAGTTAGCGAACGCTATATTCAAGCTCACATACCAA AACAAAGTGGTCAAAGTTCAACGACCGACTCCAACGGGCACGGTA ATGGATATTATATCTAGGAAAGACCAAAGGGGCAGTGGACAACTG GGAACTTATGGCCTGAATACATTCACCAACATGGAAGCCCAGTTA GTCAGACAAATGGAAGGAGAAGGTGTGCTGACAAAGGCAGACCT CGAGAACCCTCATCTGCTAGAGAAGAAAATCACACAATGGTTGGA AACCAAAGGAGTGGAGAGGTTAAAAAGAATGGCCATTAGCGGGG ATGATTGCGTGGTGAAACCAATCGATGACAGGTTCGCTAATGCCC TGCTTGCTTTGAACGATATGGGAAAGGTTCGGAAAGACATACCTC AATGGCAGCCATCAAAGGGATGGCATGATTGGCAACAGGTTCCTT TCTGCTCCCACCACTTTCATGAATTGATCATGAAAGATGGAAGAA AGTTGGTGGTTCCCTGCAGACCCCAGGACGAACTAATAGGAAGAG CAAGAATCTCTCAAGGAGCGGGATGGAGCCTTAGAGAAACTGCAT GTCTGGGGAAAGCCTACGCCCAAATGTGGAGTCTCATGTATTTTCA CAGAAGAGATCTCAGATTAGCATCCAACGCCATATGTTCAGCAGT ACCAGTCCACTGGGTTCCCACAAGTAGAACGACATGGTCTATTCAT GCTCACCATCAGTGGATGACTACAGAAGACATGCTTACTGTTTGG AACAGGGTGTGGATAGAGGAAAATCCATGGATGGAAGACAAAAC TCCAGTTACAACTTGGGAAAATGTTCCATATCTAGGAAAGAGAGA AGACCAATGGTGTGGATCACTTATTGGTCTCACTTCCAGAGCAACC TGGGCCCAGAACATACCCACAGCAATTCAACAGGTGAGAAGCCTT ATAGGCAATGAAGAGTTCCTGGACTACATGCCTTCAATGAAGAGA TTCAGGAAGGAAGAGGAGTCGGAGGGAGCCATTTGGTAAACGTA GGAAGTGGAAAAGAGGCTAACTGTCAGGCCACCTTAAGCCACAGT ACGGAAGAAGCTGTGCTGCCTGTGAGCCCCGTCCAAGGACGTTAA AAGAAGAAGTCAGGCCCCAAAGCCACGGTTTGAGCAAACCGTGCT GCCTGTAGCTCCGTCGTGGGGACGTAAAACCTGGGAGGCTGCAAA CTGTGGAAGCTGTACGCACGGTGTAGCAGACTAGCGGTTAGAGGA GACCCCTCCCATGACACAACGCAGCAGCGGGGCCCGAGCACTGAG GGAAGCTGTACCTCCTTGCAAAGGACTAGAGGTTAGAGGAGACCC CCCGCAAATAAAAACAGCATATTGACGCTGGGAGAGACCAGAGAT CCTGCTGTCTCCTCAGCATCATTCCAGGCACAGAACGCCAGAAAA TGGAATGGTGCTGTTGAATCAACAGGTTCT DEN-4 AGTTGTTAGTCTGTGTGGACCGACAAGGACAGTTCCAAATCGGAA 226 (NC_002640.1) GCTTGCTTAACACAGTTCTAACAGTTTGTTTGAATAGAGAGCAGAT CTCTGGAAAAATGAACCAACGAAAAAAGGTGGTTAGACCACCTTT CAATATGCTGAAACGCGAGAGAAACCGCGTATCAACCCCTCAAGG GTTGGTGAAGAGATTCTCAACCGGACTTTTTTCTGGGAAAGGACCC TTACGGATGGTGCTAGCATTCATCACGTTTTTGCGAGTCCTTTCCA TCCCACCAACAGCAGGGATTCTGAAGAGATGGGGACAGTTGAAGA AAAATAAGGCCATCAAGATACTGATTGGATTCAGGAAGGAGATAG GCCGCATGCTGAACATCTTGAACGGGAGAAAAAGGTCAACGATAA CATTGCTGTGCTTGATTCCCACCGTAATGGCGTTTTCCCTCAGCAC AAGAGATGGCGAACCCCTCATGATAGTGGCAAAACATGAAAGGG GGAGACCTCTCTTGTTTAAGACAACAGAGGGGATCAACAAATGCA CTCTCATTGCCATGGACTTGGGTGAAATGTGTGAGGACACTGTCAC GTATAAATGCCCCCTACTGGTCAATACCGAACCTGAAGACATTGA TTGCTGGTGCAACCTCACGTCTACCTGGGTCATGTATGGGACATGC ACCCAGAGCGGAGAACGGAGACGAGAGAAGCGCTCAGTAGCTTT AACACCACATTCAGGAATGGGATTGGAAACAAGAGCTGAGACATG GATGTCATCGGAAGGGGCTTGGAAGCATGCTCAGAGAGTAGAGAG CTGGATACTCAGAAACCCAGGATTCGCGCTCTTGGCAGGATTTATG GCTTATATGATTGGGCAAACAGGAATCCAGCGAACTGTCTTCTTTG TCCTAATGATGCTGGTCGCCCCATCCTACGGAATGCGATGCGTAGG AGTAGGAAACAGAGACTTTGTGGAAGGAGTCTCAGGTGGAGCATG GGTCGACCTGGTGCTAGAACATGGAGGATGCGTCACAACCATGGC CCAGGGAAAACCAACCTTGGATTTTGAACTGACTAAGACAACAGC CAAGGAAGTGGCTCTGTTAAGAACCTATTGCATTGAAGCCTCAAT ATCAAACATAACTACGGCAACAAGATGTCCAACGCAAGGAGAGCC TTATCTGAAAGAGGAACAGGACCAACAGTACATTTGCCGGAGAGA TGTGGTAGACAGAGGGTGGGGCAATGGCTGTGGCTTGTTTGGAAA AGGAGGAGTTGTGACATGTGCGAAGTTTTCATGTTCGGGGAAGAT AACAGGCAATTTGGTCCAAATTGAGAACCTTGAATACACAGTGGT TGTAACAGTCCACAATGGAGACACCCATGCAGTAGGAAATGACAC ATCCAATCATGGAGTTACAGCCATGATAACTCCCAGGTCACCATC GGTGGAAGTCAAATTGCCGGACTATGGAGAACTAACACTCGATTG TGAACCCAGGTCTGGAATTGACTTTAATGAGATGATTCTGATGAA AATGAAAAAGAAAACATGGCTCGTGCATAAGCAATGGTTTTTGGA TCTGCCTCTTCCATGGACAGCAGGAGCAGACACATCAGAGGTTCA CTGGAATTACAAAGAGAGAATGGTGACATTTAAGGTTCCTCATGC CAAGAGACAGGATGTGACAGTGCTGGGATCTCAGGAAGGAGCCAT GCATTCTGCCCTCGCTGGAGCCACAGAAGTGGACTCCGGTGATGG AAATCACATGTTTGCAGGACATCTTAAGTGCAAAGTCCGTATGGA GAAATTGAGAATCAAGGGAATGTCATACACGATGTGTTCAGGAAA GTTTTCAATTGACAAAGAGATGGCAGAAACACAGCATGGGACAAC AGTGGTGAAAGTCAAGTATGAAGGTGCTGGAGCTCCGTGTAAAGT CCCCATAGAGATAAGAGATGTAAACAAGGAAAAAGTGGTTGGGC GTATCATCTCATCCACCCCTTTGGCTGAGAATACCAACAGTGTAAC CAACATAGAATTAGAACCCCCCTTTGGGGACAGCTACATAGTGAT AGGTGTTGGAAACAGCGCATTAACACTCCATTGGTTCAGGAAAGG GAGTTCCATTGGCAAGATGTTTGAGTCCACATACAGAGGTGCAAA ACGAATGGCCATTCTAGGTGAAACAGCTTGGGATTTTGGTTCCGTT GGTGGACTGTTCACATCATTGGGAAAGGCTGTGCACCAGGTTTTTG GAAGTGTGTATACAACCATGTTTGGAGGAGTCTCATGGATGATTA GAATCCTAATTGGGTTCTTAGTGTTGTGGATTGGCACGAACTCGAG GAACACTTCAATGGCTATGACGTGCATAGCTGTTGGAGGAATCAC TCTGTTTCTGGGCTTCACAGTTCAAGCAGACATGGGTTGTGTGGCG TCATGGAGTGGGAAAGAATTGAAGTGTGGAAGCGGAATTTTTGTG GTTGACAACGTGCACACTTGGACAGAACAGTACAAATTTCAACCA GAGTCCCCAGCGAGACTAGCGTCTGCAATATTAAATGCCCACAAA GATGGGGTCTGTGGAATTAGATCAACCACGAGGCTGGAAAATGTC ATGTGGAAGCAAATAACCAACGAGCTAAACTATGTTCTCTGGGAA GGAGGACATGACCTCACTGTAGTGGCTGGGGATGTGAAGGGGGTG TTGACCAAAGGCAAGAGAGCACTCACACCCCCAGTGAGTGATCTG AAATATTCATGGAAGACATGGGGAAAAGCAAAAATCTTCACCCCA GAAGCAAGAAATAGCACATTTTTAATAGACGGACCAGACACCTCT GAATGCCCCAATGAACGAAGAGCATGGAACTCTCTTGAGGTGGAA GACTATGGATTTGGCATGTTCACGACCAACATATGGATGAAATTCC GAGAAGGAAGTTCAGAAGTGTGTGACCACAGGTTAATGTCAGCTG CAATTAAAGATCAGAAAGCTGTGCATGCTGACATGGGTTATTGGA TAGAGAGCTCAAAAAACCAGACCTGGCAGATAGAGAAAGCATCTC TTATTGAAGTGAAAACATGTCTGTGGCCCAAGACCCACACACTGT GGAGCAATGGAGTGCTGGAAAGCCAGATGCTCATTCCAAAATCAT ATGCGGGCCCTTTTTCACAGCACAATTACCGCCAGGGCTATGCCAC GCAAACCGTGGGCCCATGGCACTTAGGCAAATTAGAGATAGACTT TGGAGAATGCCCCGGAACAACAGTCACAATTCAGGAGGATTGTGA CCATAGAGGCCCATCTTTGAGGACCACCACTGCATCTGGAAAACT AGTCACGCAATGGTGCTGCCGCTCCTGCACGATGCCTCCCTTAAGG TTCTTGGGAGAAGATGGGTGCTGGTATGGGATGGAGATTAGGCCC TTGAGTGAAAAAGAAGAGAACATGGTCAAATCACAGGTGACGGC CGGACAGGGCACATCAGAAACTTTTTCTATGGGTCTGTTGTGCCTG ACCTTGTTTGTGGAAGAATGCTTGAGGAGAAGAGTCACTAGGAAA CACATGATATTAGTTGTGGTGATCACTCTTTGTGCTATCATCCTGG GAGGCCTCACATGGATGGACTTACTACGAGCCCTCATCATGTTGG GGGACACTATGTCTGGTAGAATAGGAGGACAGATCCACCTAGCCA TCATGGCAGTGTTCAAGATGTCACCAGGATACGTGCTGGGTGTGTT TTTAAGGAAACTCACTTCAAGAGAGACAGCACTAATGGTAATAGG AATGGCCATGACAACGGTGCTTTCAATTCCACATGACCTTATGGAA CTCATTGATGGAATATCACTGGGACTAATTTTGCTAAAAATAGTAA CACAGTTTGACAACACCCAAGTGGGAACCTTAGCTCTTTCCTTGAC TTTCATAAGATCAACAATGCCATTGGTCATGGCTTGGAGGACCATT ATGGCTGTGTTGTTTGTGGTCACACTCATTCCTTTGTGCAGGACAA GCTGTCTTCAAAAACAGTCTCATTGGGTAGAAATAACAGCACTCA TCCTAGGAGCCCAAGCTCTGCCAGTGTACCTAATGACTCTTATGAA AGGAGCCTCAAGAAGATCTTGGCCTCTTAACGAGGGCATAATGGC TGTGGGTTTGGTTAGTCTCTTAGGAAGCGCTCTTTTAAAGAATGAT GTCCCTTTAGCTGGCCCAATGGTGGCAGGAGGCTTACTTCTGGCGG CTTACGTGATGAGTGGTAGCTCAGCAGATCTGTCACTAGAGAAGG CCGCCAACGTGCAGTGGGATGAAATGGCAGACATAACAGGCTCAA GCCCAATCGTAGAAGTGAAGCAGGATGAAGATGGCTCTTTCTCCA TACGGGACGTCGAGGAAACCAATATGATAACCCTTTTGGTGAAAC TGGCACTGATAACAGTGTCAGGTCTCTACCCCTTGGCAATTCCAGT CACAATGACCTTATGGTACATGTGGCAAGTGAAAACACAAAGATC AGGAGCCCTGTGGGACGTCCCCTCACCCGCTGCCACTAAAAAAGC CGCACTGTCTGAAGGAGTGTACAGGATCATGCAAAGAGGGTTATT CGGGAAAACTCAGGTTGGAGTAGGGATACACATGGAAGGTGTATT TCACACAATGTGGCATGTAACAAGAGGATCAGTGATCTGCCACGA GACTGGGAGATTGGAGCCATCTTGGGCTGACGTCAGGAATGACAT GATATCATACGGTGGGGGATGGAGGCTTGGAGACAAATGGGACA AAGAAGAAGACGTTCAGGTCCTCGCCATAGAACCAGGAAAAAATC CTAAACATGTCCAAACGAAACCTGGCCTTTTCAAGACCCTAACTG GAGAAATTGGAGCAGTAACATTAGATTTCAAACCCGGAACGTCTG GTTCTCCCATCATCAACAGGAAAGGAAAAGTCATCGGACTCTATG GAAATGGAGTAGTTACCAAATCAGGTGATTACGTCAGTGCCATAA CGCAAGCCGAAAGAATTGGAGAGCCAGATTATGAAGTGGATGAG GACATTTTTCGAAAGAAAAGATTAACTATAATGGACTTACACCCC GGAGCTGGAAAGACAAAAAGAATTCTTCCATCAATAGTGAGAGAA GCCTTAAAAAGGAGGCTACGAACTTTGATTTTAGCTCCCACGAGA GTGGTGGCGGCCGAGATGGAAGAGGCCCTACGTGGACTGCCAATC CGTTATCAGACCCCAGCTGTGAAATCAGAACACACAGGAAGAGAG ATTGTAGACCTCATGTGTCATGCAACCTTCACAACAAGACTTTTGT CATCAACCAGGGTTCCAAATTACAACCTTATAGTGATGGATGAAG CACATTTCACCGATCCTTCTAGTGTCGCGGCTAGAGGATACATCTC GACCAGGGTGGAAATGGGAGAGGCAGCAGCCATCTTCATGACCGC AACCCCTCCCGGAGCGACAGATCCCTTTCCCCAGAGCAACAGCCC AATAGAAGACATCGAGAGGGAAATTCCGGAAAGGTCATGGAACA CAGGGTTCGACTGGATAACAGACTACCAAGGGAAAACTGTGTGGT TTGTTCCCAGCATAAAAGCTGGAAATGACATTGCAAATTGTTTGAG AAAGTCGGGAAAGAAAGTTATCCAGTTGAGTAGGAAAACCTTTGA TACAGAGTATCCAAAAACGAAACTCACGGACTGGGACTTTGTGGT CACTACAGACATATCTGAAATGGGGGCCAATTTTAGAGCCGGGAG AGTGATAGACCCTAGAAGATGCCTCAAGCCAGTTATCCTACCAGA TGGGCCAGAGAGAGTCATTTTAGCAGGTCCTATTCCAGTGACTCCA GCAAGCGCTGCTCAGAGAAGAGGGCGAATAGGAAGGAACCCAGC ACAAGAAGACGACCAATACGTTTTCTCCGGAGACCCACTAAAAAA TGATGAAGATCATGCCCACTGGACAGAAGCAAAGATGCTGCTTGA CAATATCTACACCCCAGAAGGGATCATTCCAACATTGTTTGGTCCG GAAAGGGAAAAAACCCAAGCCATTGATGGAGAGTTTCGCCTCAGA GGGGAACAAAGGAAGACTTTTGTGGAATTAATGAGGAGAGGAGA CCTTCCGGTGTGGCTGAGCTATAAGGTAGCTTCTGCTGGCATTTCT TACGAAGATCGGGAATGGTGCTTCACAGGGGAAAGAAATAACCA AATTTTAGAAGAAAACATGGAGGTTGAAATTTGGACTAGAGAGGG AGAAAAGAAAAAGCTAAGGCCAAGATGGTTAGATGCACGTGTAT ACGCTGACCCCATGGCTTTGAAGGATTTCAAGGAGTTTGCCAGTG GAAGGAAGAGTATAACTCTCGACATCCTAACAGAGATTGCCAGTT TGCCAACTTACCTTTCCTCTAGGGCCAAGCTCGCCCTTGATAACAT AGTCATGCTCCACACAACAGAAAGAGGAGGGAGGGCCTATCAAC ACGCCCTGAACGAACTTCCGGAGTCACTGGAAACACTCATGCTTG TAGCTTTACTAGGTGCTATGACAGCAGGCATCTTCCTGTTTTTCAT GCAAGGGAAAGGAATAGGGAAATTGTCAATGGGTTTGATAACCAT TGCGGTGGCTAGTGGCTTGCTCTGGGTAGCAGAAATTCAACCCCA GTGGATAGCGGCCTCAATCATACTAGAGTTTTTTCTCATGGTACTG TTGATACCGGAACCAGAAAAACAAAGGACCCCACAAGACAATCA ATTGATCTACGTCATATTGACCATTCTCACCATCATTGGTCTAATA GCAGCCAACGAGATGGGGCTGATTGAAAAAACAAAAACGGATTTT GGGTTTTACCAGGTAAAAACAGAAACCACCATCCTCGATGTGGAC TTGAGACCAGCTTCAGCATGGACGCTCTATGCAGTAGCCACCACA ATTCTGACTCCCATGCTGAGACACACCATAGAAAACACGTCGGCC AACCTATCTCTAGCAGCCATTGCCAACCAGGCAGCCGTCCTAATG GGGCTTGGAAAAGGATGGCCGCTCCACAGAATGGACCTCGGTGTG CCGCTGTTAGCAATGGGATGCTATTCTCAAGTGAACCCAACAACCT TGACAGCATCCTTAGTCATGCTTTTAGTCCATTATGCAATAATAGG CCCAGGATTGCAGGCAAAAGCCACAAGAGAGGCCCAGAAAAGGA CAGCTGCTGGGATCATGAAAAATCCCACAGTGGACGGGATAACAG TAATAGATCTAGAACCAATATCCTATGACCCAAAATTTGAAAAGC AATTAGGGCAGGTCATGCTACTAGTCTTGTGTGCTGGACAACTACT CTTGATGAGAACAACATGGGCTTTCTGTGAAGTCTTGACTTTGGCC ACAGGACCAATCTTGACCTTGTGGGAGGGCAACCCGGGAAGGTTT TGGAACACGACCATAGCCGTATCCACCGCCAACATTTTCAGGGGA AGTTACTTGGCGGGAGCTGGACTGGCTTTTTCACTCATAAAGAATG CACAAACCCCTAGGAGGGGAACTGGGACCACAGGAGAGACACTG GGAGAGAAGTGGAAGAGACAGCTAAACTCATTAGACAGAAAAGA GTTTGAAGAGTATAAAAGAAGTGGAATACTAGAAGTGGACAGGA CTGAAGCCAAGTCTGCCCTGAAAGATGGGTCTAAAATCAAGCATG CAGTATCAAGAGGGTCCAGTAAGATCAGATGGATTGTTGAGAGAG GGATGGTAAAGCCAAAAGGGAAAGTTGTAGATCTTGGCTGTGGGA GAGGAGGATGGTCTTATTACATGGCGACACTCAAGAACGTGACTG AAGTGAAAGGGTATACAAAAGGAGGTCCAGGACATGAAGAACCG ATTCCCATGGCTACTTATGGTTGGAATTTGGTCAAACTCCATTCAG GGGTTGACGTGTTCTACAAACCCACAGAGCAAGTGGACACCCTGC TCTGTGATATTGGGGAGTCATCTTCTAATCCAACAATAGAGGAAG GAAGAACATTAAGAGTTTTGAAGATGGTGGAGCCATGGCTCTCTT CAAAACCTGAATTCTGCATCAAAGTCCTTAACCCCTACATGCCAAC AGTCATAGAAGAGCTGGAGAAACTGCAGAGAAAACATGGTGGGA ACCTTGTCAGATGCCCGCTGTCCAGGAACTCCACCCATGAGATGTA TTGGGTGTCAGGAGCGTCGGGAAACATTGTGAGCTCTGTGAACAC AACATCAAAGATGTTGTTGAACAGGTTCACAACAAGGCATAGGAA ACCCACTTATGAGAAGGACGTAGATCTTGGGGCAGGAACGAGAAG TGTCTCCACTGAAACAGAAAAACCAGACATGACAATCATTGGGAG AAGGCTTCAGCGATTGCAAGAAGAGCACAAAGAAACCTGGCATTA TGATCAGGAAAACCCATACAGAACCTGGGCGTATCATGGAAGCTA TGAAGCTCCTTCGACAGGCTCTGCATCCTCCATGGTGAACGGGGTG GTAAAACTGCTAACAAAACCCTGGGATGTGATTCCAATGGTGACT CAGTTAGCCATGACAGATACAACCCCTTTTGGGCAACAAAGAGTG TTCAAAGAGAAGGTGGATACCAGAACACCACAACCAAAACCCGGT ACACGAATGGTTATGACCACGACAGCCAATTGGCTGTGGGCCCTC CTTGGAAAGAAGAAAAATCCCAGACTGTGCACAAGGGAAGAGTTC ATCTCAAAAGTTAGATCAAACGCAGCCATAGGCGCAGTCTTTCAG GAAGAACAGGGATGGACATCAGCCAGTGAAGCTGTGAATGACAG CCGGTTTTGGGAACTGGTTGACAAAGAAAGGGCCCTACACCAGGA AGGGAAATGTGAATCGTGTGTCTATAACATGATGGGAAAACGTGA GAAAAAGTTAGGAGAGTTTGGCAGAGCCAAGGGAAGCCGAGCAA TCTGGTACATGTGGCTGGGAGCGCGGTTTCTGGAATTTGAAGCCCT GGGTTTTTTGAATGAAGATCACTGGTTTGGCAGAGAAAATTCATG GAGTGGAGTGGAAGGGGAAGGTCTGCACAGATTGGGATATATCCT GGAGGAGATAGACAAGAAGGATGGAGACCTAATGTATGCTGATG ACACAGCAGGCTGGGACACAAGAATCACTGAGGATGACCTTCAAA ATGAGGAACTGATCACGGAACAGATGGCTCCCCACCACAAGATCC TAGCCAAAGCCATTTTCAAACTAACCTATCAAAACAAAGTGGTGA AAGTCCTCAGACCCACACCGCGGGGAGCGGTGATGGATATCATAT CCAGGAAAGACCAAAGAGGTAGTGGACAAGTTGGAACATATGGTT TGAACACATTCACCAACATGGAAGTTCAACTCATCCGCCAAATGG AAGCTGAAGGAGTCATCACACAAGATGACATGCAGAACCCAAAA GGGTTGAAAGAAAGAGTTGAGAAATGGCTGAAAGAGTGTGGTGTC GACAGGTTAAAGAGGATGGCAATCAGTGGAGACGATTGCGTGGTG AAGCCCCTAGATGAGAGGTTTGGCACTTCCCTCCTCTTCTTGAACG ACATGGGAAAGGTGAGGAAAGACATTCCGCAGTGGGAACCATCTA AGGGATGGAAAAACTGGCAAGAGGTTCCTTTTTGCTCCCACCACTT TCACAAGATCTTTATGAAGGATGGCCGCTCACTAGTTGTTCCATGT AGAAACCAGGATGAACTGATAGGGAGAGCCAGAATCTCGCAGGG AGCTGGATGGAGCTTAAGAGAAACAGCCTGCCTGGGCAAAGCTTA CGCCCAGATGTGGTCGCTTATGTACTTCCACAGAAGGGATCTGCGT TTAGCCTCCATGGCCATATGCTCAGCAGTTCCAACGGAATGGTTTC CAACAAGCAGAACAACATGGTCAATCCACGCTCATCACCAGTGGA TGACCACTGAAGATATGCTCAAAGTGTGGAACAGAGTGTGGATAG AAGACAACCCTAATATGACTGACAAGACTCCAGTCCATTCGTGGG AAGATATACCTTACCTAGGGAAAAGAGAGGATTTGTGGTGTGGAT CCCTGATTGGACTTTCTTCCAGAGCCACCTGGGCGAAGAACATTCA TACGGCCATAACCCAGGTCAGGAACCTGATCGGAAAAGAGGAATA CGTGGATTACATGCCAGTAATGAAAAGATACAGTGCTCCTTCAGA GAGTGAAGGAGTTCTGTAATTACCAACAACAAACACCAAAGGCTA TTGAAGTCAGGCCACTTGTGCCACGGTTTGAGCAAACCGTGCTGCC TGTAGCTCCGCCAATAATGGGAGGCGTAATAATCCCCAGGGAGGC CATGCGCCACGGAAGCTGTACGCGTGGCATATTGGACTAGCGGTT AGAGGAGACCCCTCCCATCACTGATAAAACGCAGCAAAAGGGGG CCCGAAGCCAGGAGGAAGCTGTACTCCTGGTGGAAGGACTAGAGG TTAGAGGAGACCCCCCCAACACAAAAACAGCATATTGACGCTGGG AAAGACCAGAGATCCTGCTGTCTCTGCAACATCAATCCAGGCACA GAGCGCCGCAAGATGGATTGGTGTTGTTGATCCAACAGGTTCT Construct 1 ATGGATGCTATGAAAAGAGGCCTGTGTTGTGTGTTGCTGTTGTGCG 227 GAGCTGTGTTTGTGTCACCTTTCCACCTGACTACCCGCAATGGTGA GCCCCATATGATTGTGTCGCGCCAGGAGAAGGGGAAGTCCCTCCT GTTCAAAACTGAAAACGGCGTGAACATGTGTACCCTGATGGCCAT GGACCTTGGAGAACTGTGCGAGGACACCATCACCTACAATTGTCC GCTCCTGCGCCAAAACGAACCAGAAGATATCGACTGCTGGTGCAA TTCCACTTCAACCTGGGTTACCTACGGAACTTGCACCGCCACGGGA GAACACAGAAGAGAAAAGCGCTCGGTGGCGCTGGTGCCTCATGTC GGAATGGGACTGGAGACTCGGACGGAGACTTGGATGTCCTCGGAG GGAGCATGGAAACATGCCCAACGGATCGAAACTTGGGTGCTGAGG CACCCTGGATTCACCATCATGGCAGCGATCCTCGCCTACACTATAG GTACTACCTACTTTCAAAGGGTGCTGATCTTCATTCTCCTCACCGC AGTGGCCCCTTCAATGACCATGAGGTGCATTGGGATCTCGAACCG GGACTTCGTCGAAGGAGTGTCCGGAGGTAGCTGGGTCGACATCGT CCTGGAACACGGAAGCTGCGTGACTACTATGGCGAAGAACAAGCC AACCTTGGACTTCGAGCTTATCAAGACCGAGGCGAAGCACCCGGC CACTCTGAGAAAGTACTGCATCGAGGCTAAGCTCACCAACACGAC CACTGCCTCGCGATGCCCAACTCAGGGAGAACCGTCACTGAACGA AGAACAGGATAAACGCTTTGTGTGCAAGCATAGCATGGTGGATAG AGGCTGGGGAAACGGCTGTGGACTCTTCGGAAAGGGTGGAATTGT GACGTGCGCAATGTTCACTTGCAAGAAGAATATGGAAGGGAAGAT CGTCCAGCCGGAGAACCTGGAATACACTATCGTGATCACCCCGCA CTCAGGCGAGGAGAACGCAGTGGGCAACGATACCGGGAAGCACG GGAAGGAAATCAAGGTGACCCCGCAGTCGTCCATTACCGAGGCCG AACTCACCGGATACGGCACTGTGACTATGGAATGCTCGCCACGGA CCGGGCTGGATTTCAATGAGATGGTGCTCTTGCAAATGGAGAACA AAGCCTGGCTGGTCCACCGCCAGTGGTTCCTCGACCTCCCCCTTCC GTGGCTGCCGGGAGCTGACACCCAAGGATCCAACTGGATCCAAAA AGAAACCCTTGTCACGTTTAAGAATCCACATGCCAAAAAGCAGGA CGTGGTCGTGCTCGGAAGCCAGGAAGGAGCCATGCACACTGCGCT GACTGGAGCAACCGAAATTCAAATGTCGAGCGGCAACCTCCTCTT CACTGGACATCTGAAGTGCCGGCTGCGCATGGACAAACTGCAACT TAAGGGCATGTCATACTCGATGTGTACCGGCAAATTCAAGGTGGT GAAGGAGATCGCGGAGACTCAGCACGGGACCATCGTCATCCGGGT CCAGTATGAGGGTGATGGTTCCCCCTGCAAGATCCCTTTCGAAATC ATGGATCTGGAGAAACGTCACGTGCTGGGCCGGCTGATCACTGTG AATCCGATCGTTACGGAGAAAGACAGCCCGGTGAACATCGAAGCT GAACCGCCGTTTGGGGATAGCTACATTATCATCGGCGTGGAACCA GGCCAGCTCAAGTTGTCGTGGTTCAAGAAAGGATCCAGCATCGGA CAGATGTTCGAAACCACTATGCGCGGAGCCAAACGCATGGCTATC CTGGGGGACACGGCCTGGGACTTCGGGTCGCTGGGTGGTGTGTTC ACCTCCATTGGAAAGGCGCTCCATCAGGTGTTTGGTGCGATCTACG GCGCCGCATTCTCCGGAGTGTCATGGACCATGAAGATCCTCATCG GAGTCGTCATCACCTGGATCGGCATGAATTCTCGGTCCACTTCCTT GAGCGTCAGCCTGGTGCTGGTCGGAGTTGTGACTCTGTACCTTGGA GTGATGGTCCAGGCC Construct 2 ATGGATGCTATGAAAAGAGGCCTGTGTTGTGTGTTGCTGTTGTGCG 228 GAGCTGTGTTTGTGTCACCTTTCCACCTGACTACCCGCAATGGTGA GCCCCATATGATTGTGTCGCGCCAGGAGAAGGGGAAGTCCCTCCT GTTCAAAACTGAAAACGGCGTGAACATGTGTACCCTGATGGCCAT GGACCTTGGAGAACTGTGCGAGGACACCATCACCTACAATTGTCC GCTCCTGCGCCAAAACGAACCAGAAGATATCGACTGCTGGTGCAA TTCCACTTCAACCTGGGTTACCTACGGAACTTGCACCGCCACGGGA GAACACAGAAGAGAAAAGCGCTCGGTGGCGCTGGTGCCTCATGTC GGAATGGGACTGGAGACTCGGACGGAGACTTGGATGTCCTCGGAG GGAGCATGGAAACATGCCCAACGGATCGAAACTTGGGTGCTGAGG CACCCTGGATTCACCATCATGGCAGCGATCCTCGCCTACACTATAG GTACTACCTACTTTCAAAGGGTGCTGATCTTCATTCTCCTCACCGC AGTGGCCCCTTCAATGACCATGAGGTGCATTGGGATCTCGAACCG GGACTTCGTCGAAGGAGTGTCCGGAGGTAGCTGGGTCGACATCGT CCTGGAACACGGAAGCTGCGTGACTACTATGGCGAAGAACAAGCC AACCTTGGACTTCGAGCTTATCAAGACCGAGGCGAAGCACCCGGC CACTCTGAGAAAGTACTGCATCGAGGCTAAGCTCACCAACACGAC CACTGCCTCGCGATGCCCAACTCAGGGAGAACCGTCACTGAACGA AGAACAGGATAAACGCTTTGTGTGCAAGCATAGCATGGTGGATAG AGGCTGGGGAAACGGCTGTGGACTCTTCGGAAAGGGTGGAATTGT GACGTGCGCAATGTTCACTTGCAAGAAGAATATGGAAGGGAAGAT CGTCCAGCCGGAGAACCTGGAATACACTATCGTGATCACCCCGCA CTCAGGCGAGGAGAACGCAGTGGGCAACGATACCGGGAAGCACG GGAAGGAAATCAAGGTGACCCCGCAGTCGTCCATTACCGAGGCCG AACTCACCGGATACGGCACTGTGACTATGGAATGCTCGCCACGGA CCGGGCTGGATTTCAATGAGATGGTGCTCTTGCAAATGGAGAACA AAGCCTGGCTGGTCCACCGCCAGTGGTTCCTCGACCTCCCCCTTCC GTGGCTGCCGGGAGCTGACACCCAAGGATCCAACTGGATCCAAAA AGAAACCCTTGTCACGTTTAAGAATCCACATGCCAAAAAGCAGGA CGTGGTCGTGCTCGGAAGCCAGGAAGGAGCCATGCACACTGCGCT GACTGGAGCAACCGAAATTCAAATGTCGAGCGGCAACCTCCTCTT CACTGGACATCTGAAGTGCCGGCTGCGCATGGACAAACTGCAACT TAAGGGCATGTCATACTCGATGTGTACCGGCAAATTCAAGGTGGT GAAGGAGATCGCGGAGACTCAGCACGGGACCATCGTCATCCGGGT CCAGTATGAGGGTGATGGTTCCCCCTGCAAGATCCCTTTCGAAATC ATGGATCTGGAGAAACGTCACGTGCTGGGCCGGCTGATCACTGTG AATCCGATCGTTACGGAGAAAGACAGCCCGGTGAACATCGAAGCT GAACCGCCGTTTGGGGATAGCTACATTATCATCGGCGTGGAACCA GGCCAGCTCAAGTTGTCGTGGTTCAAGAAAGGA Construct 3 ATGGATGCTATGAAAAGAGGCCTGTGTTGTGTGTTGCTGTTGTGCG 229 GAGCTGTGTTTGTGTCACCTTTCCACCTGACTACCCGCAATGGTGA GCCCCATATGATTGTGTCGCGCCAGGAGAAGGGGAAGTCCCTCCT GTTCAAAACTGAAAACGGCGTGAACATGTGTACCCTGATGGCCAT GGACCTTGGAGAACTGTGCGAGGACACCATCACCTACAATTGTCC GCTCCTGCGCCAAAACGAACCAGAAGATATCGACTGCTGGTGCAA TTCCACTTCAACCTGGGTTACCTACGGAACTTGCACCGCCACGGGA GAACACAGAAGAGAAAAGCGCTCGGTGGCGCTGGTGCCTCATGTC GGAATGGGACTGGAGACTCGGACGGAGACTTGGATGTCCTCGGAG GGAGCATGGAAACATGCCCAACGGATCGAAACTTGGGTGCTGAGG CACCCTGGATTCACCATCATGGCAGCGATCCTCGCCTACACTATAG GTACTACCTACTTTCAAAGGGTGCTGATCTTCATTCTCCTCACCGC AGTGGCCCCTTCAATGACCATGAGGTGCATTGGGATCTCGAACCG GGACTTCGTCGAAGGAGTGTCCGGAGGTAGCTGGGTCGACATCGT CCTGGAACACGGAAGCTGCGTGACTACTATGGCGAAGAACAAGCC AACCTTGGACTTCGAGCTTATCAAGACCGAGGCGAAGCACCCGGC CACTCTGAGAAAGTACTGCATCGAGGCTAAGCTCACCAACACGAC CACTGCCTCGCGATGCCCAACTCAGGGAGAACCGTCACTGAACGA AGAACAGGATAAACGCTTTGTGTGCAAGCATAGCATGGTGGATAG AGGCTGGGGAAACGGCTGTGGACTCTTCGGAAAGGGTGGAATTGT GACGTGCGCAATGTTCACTTGCAAGAAGAATATGGAAGGGAAGAT CGTCCAGCCGGAGAACCTGGAATACACTATCGTGATCACCCCGCA CTCAGGCGAGGAGAACGCAGTGGGCAACGATACCGGGAAGCACG GGAAGGAAATCAAGGTGACCCCGCAGTCGTCCATTACCGAGGCCG AACTCACCGGATACGGCACTGTGACTATGGAATGCTCGCCACGGA CCGGGCTGGATTTCAATGAGATGGTGCTCTTGCAAATGGAGAACA AAGCCTGGCTGGTCCACCGCCAGTGGTTCCTCGACCTCCCCCTTCC GTGGCTGCCGGGAGCTGACACCCAAGGATCCAACTGGATCCAAAA AGAAACCCTTGTCACGTTTAAGAATCCACATGCCAAAAAGCAGGA CGTGGTCGTGCTCGGAAGCCAGGAAGGAGCCATGCACACTGCGCT GACTGGAGCAACCGAAATTCAAATGTCGAGCGGCAACCTCCTCTT CACTGGACATCTGAAGTGCCGGCTGCGCATGGACAAACTGCAACT TAAGGGCATGTCATACTCGATGTGTACCGGCAAATTCAAGGTGGT GAAGGAGATCGCGGAGACTCAGCACGGGACCATCGTCATCCGGGT CCAGTATGAGGGTGATGGTTCCCCCTGCAAGATCCCTTTCGAAATC ATGGATCTGGAGAAACGTCACGTGCTGGGCCGGCTGATCACTGTG AATCCGATCGTTACGGAGAAAGACAGCCCGGTGAACATCGAAGCT GAACCGCCGTTTGGGGATAGCTACATTATCATCGGCGTGGAACCA GGCCAGCTCAAGTTGTCGTGGTTCAAGAAAGGAGGAGGTGGAGGA TCCGGAGGCGGAGGGTCGGGCGGTGGTGGATCGGAGGTCAAACTG CAGCAATCAGGGACCGAAGTCGTGAAGCCGGGGGCTTCAGTCAAG CTGTCCTGCAAGGCCAGCGGCTATATCTTCACTAGCTACGACATCG ATTGGGTGCGGCAGACTCCGGAGCAAGGACTCGAGTGGATTGGGT GGATCTTTCCGGGCGAGGGATCAACCGAGTACAACGAAAAATTTA AGGGACGGGCAACGCTGTCCGTGGACAAGAGCTCATCTACGGCGT ACATGGAGCTGACGCGGCTCACGTCAGAGGATTCCGCCGTCTACT TCTGTGCCAGGGGCGACTACTACCGGCGCTACTTTGATCTGTGGGG ACAAGGAACGACCGTGACTGTCTCATCAGGCGGCGGCGGATCGGG AGGAGGCGGATCGGGTGGCGGTGGTTCGGACATTCAGATGACTCA ATCGCCCAGCTTCCTGTCGACCTCACTGGGGAATTCTATTACGATC ACTTGTCACGCTTCGCAGAACATCAAGGGTTGGCTGGCTTGGTACC AGCAGAAAAGCGGTAACGCCCCGCAACTGCTCATCTACAAGGCAT CGTCCCTGCAATCGGGAGTGCCGTCACGCTTTTCAGGATCGGGCTC CGGAACCGATTACATCTTTACCATCAGCAACCTGCAGCCGGAAGA CATCGCCACTTACTACTGTCAACACTATCAGAGCTTTCCGTGGACC TTTGGAGGGGGGACCAAATTGGAGATCAAGCGCGACTACAAGGAT GACGATGACAAA Construct 4 ATGGATTGGACCTGGATCTTGTTTCTCGTCGCCGCAGCCACTCGCG 230 TTCATAGCAAAGGAATGTCATACTCCATGTGCACGGGAAAATTCA AGGTGGTCAAAGAGATCGCGGAGACTCAGCACGGCACCATCGTCA TTCGCGTGCAAACTGAAGGAGATGGATCTCCCTGCAAGATCCCGT TCGAGATCATGGACCTGGAAAAGAGACACGTCCTCGGTAGACTGA TCACCGTGAACCCGATCGTGACGGAGAAGGATTCCCCGGTGAATA TTGAAGCAGAGCCTCCATTTGGGGACTCATACATTATCATTGGGGT CGAGCCGGGCCAGCTGAAGCTGAATTGGTTTAAGAAGGGCTCGTC AATCGGACAGATGTTCGAAACTACTATGAGGGGTGCAAAGCGGAT GGCGATCCTCTCGGGCGGAGATATCATCAAACTCCTTAACGAACA GGTGAACAAGGAGATGCAGTCCTCAAACCTTTACATGAGCATGTC GTCCTGGTGTTACACCCATAGCCTGGACGGCGCTGGATTGTTCCTG TTTGACCATGCAGCGGAGGAATACGAACACGCCAAGAAGCTCATC ATCTTCCTGAACGAGAATAACGTGCCAGTGCAACTGACCTCCATCT CGGCTCCTGAGCACAAGTTCGAAGGACTCACCCAGATCTTCCAAA AGGCCTACGAACACGAACAGCACATCAGCGAATCCATCAACAATA TCGTGGACCATGCTATCAAAAGCAAAGACCATGCGACCTTCAACT TCCTGCAATGGTATGTCGCCGAACAGCACGAAGAGGAGGTGCTGT TCAAGGACATTCTCGACAAAATCGAATTGATAGGGAACGAAAATC ACGGTCTGTACCTGGCCGATCAATACGTGAAGGGAATTGCCAAGT CGCGGAAGTCGT Dengue 2 prME ATGCTGAATATTCTGAACCGCCGCCGCCGGACTGCCGGGATTATA 231 (Thailand/0168/1979) ATTATGATGATTCCCACCGTGATGGCCTTCCACCTGACCACCCGGA ACGGGGAACCACATATGATCGTGTCCAGACAGGAGAAGGGAAAG TCCCTGCTGTTCAAGACCGAGGACGGCGTGAACATGTGCACCCTC ATGGCTATGGACCTGGGCGAACTCTGCGAGGACACCATCACCTAC AAGTGCCCCCTGTTGAGGCAGAACGAGCCGGAGGATATTGACTGC TGGTGCAATTCGACCAGCACCTGGGTCACCTACGGGACTTGCACC ACAACCGGAGAACATCGGCGCGAAAAGCGCAGCGTGGCTTTGGTG CCTCACGTCGGAATGGGGCTGGAGACTAGAACCGAGACTTGGATG TCGTCGGAAGGGGCCTGGAAACACGCACAGCGCATCGAAACTTGG ATACTCAGGCATCCCGGCTTCACCATTATGGCCGCGATCCTGGCAT ACACCATCGGTACTACCCACTTCCAACGGGCCCTGATCTTTATCCT CCTGACCGCTGTCGCACCATCCATGACCATGCGGTGTATCGGTATC AGCAACAGGGACTTCGTGGAGGGAGTGTCGGGAGGATCCTGGGTG GATATTGTGCTGGAACACGGTTCCTGCGTCACTACCATGGCGAAG AACAAGCCTACCCTGGACTTTGAGCTGATCAAAACTGAGGCCAAG CAGCCGGCCACCCTGCGCAAGTACTGCATCGAAGCCAAGCTGACC AATACCACTACCGAATCCCGCTGTCCGACCCAAGGGGAGCCCTCC CTGAATGAGGAGCAGGACAAGCGCTTCGTCTGCAAGCATTCAATG GTCGACCGCGGCTGGGGAAACGGCTGCGGACTGTTCGGAAAGGGC GGCATTGTGACCTGTGCCATGTTCACTTGCAAGAAGAACATGGAA GGAAAGATCGTGCAGCCCGAAAACCTGGAGTATACCATCGTCGTG ACCCCGCACTCCGGGGAAGAACACGCTGTGGGAAACGACACCGG AAAGCACGGAAAGGAGATCAAAGTGACCCCACAGTCGAGCATTA CCGAGGCCGAACTTACTGGTTACGGCACTGTGACGATGGAATGTT CACCGAGAACTGGACTGGATTTCAACGAAATGGTGCTGCTCCAAA TGGAAAACAAGGCCTGGCTGGTGCACCGCCAGTGGTTTCTTGACC TCCCTCTCCCTTGGCTGCCTGGAGCAGACACTCAGGGTTCCAACTG GATTCAGAAGGAAACACTCGTGACCTTCAAGAACCCTCACGCGAA GAAGCAGGATGTGGTCGTGCTGGGAAGCCAGGAGGGAGCGATGC ATACCGCCCTCACCGGCGCGACGGAGATTCAGATGTCCAGCGGAA ACCTTCTGTTCACCGGACACCTGAAGTGCAGACTGAGGATGGACA AGCTGCAGCTCAAGGGAATGTCCTACTCCATGTGCACTGGAAAGT TCAAGGTCGTGAAGGAGATTGCCGAAACTCAGCATGGTACCATCG TGATCCGGGTGCAATATGAAGGGGACGGATCCCCGTGCAAGATCC CTTTCGAAATCATGGACTTGGAGAAGCGACACGTGCTGGGCAGAC TGATCACAGTCAACCCCATCGTGACTGAGAAGGATTCACCCGTGA ACATTGAAGCCGAGCCGCCTTTCGGCGATAGCTACATCATCATTGG CGTGGAACCGGGACAGCTTAAGCTCAACTGGTTCAAGAAGGGTTC CTCGATCGGTCAAATGTTTGAAACCACGATGCGGGGTGCCAAACG GATGGCCATTCTGGGAGACACCGCCTGGGATTTCGGCTCCTTGGGC GGAGTGTTCACTTCTATCGGAAAGGCGCTGCACCAAGTGTTCGGA GCCATCTACGGCGCCGCGTTCTCGGGCGTCAGCTGGACCATGAAG ATTCTGATCGGGGTCATCATCACTTGGATTGGGATGAACTCACGGT CCACCTCCCTGAGCGTGTCCCTTGTCCTGGTCGGCATCGTGACCCT GTACCTCGGAGTGATGGTGCAGGCTTAG Dengue 2 prME ATGCTTAACATTCTCAACCGCCGCCGGAGAACTGCTGGTATTATCA 232 (Thailand/16681/1984) TTATGATGATTCCCACTGTGATGGCCTTCCACCTGACCACGCGGAA CGGCGAACCCCATATGATTGTCGGTCGGCAGGAAAAGGGGAAGTC CCTGCTGTTCAAAACTGAGGACGGAGTGAACATGTGCACCCTCAT GGCTATTGACCTGGGAGAGCTGTGCGAAGATACTATCACGTACAA GTGCCCCCTGCTGCGCCAGAACGAGCCTGAGGACATTGACTGCTG GTGCAACTCCACGTCAACCTGGGTCACCTACGGAACTTGCGCGAC TACCGGCGAACATCGCAGAGAAAAGAGAAGCGTGGCCCTCGTGCC GCACGTCGGGATGGGGCTGGAAACCCGGACCGAAACCTGGATGTC CTCGGAAGGCGCCTGGAAGCACGTGCAGAGGATCGAAACTTGGAT CCTCCGGCACCCGGGATTCACCATCATGGCCGCCATCCTCGCTTAC ACAATCGGAACCACTCACTTTCAACGCGCCCTGATCTTCATCCTGC TTACCGCCGTGGCCCCGTCCATGACCATGCGCTGCATTGGAATGTC AAACCGGGACTTCGTCGAGGGAGTCTCCGGAGGAAGCTGGGTGGA CATCGTGCTGGAGCACGGCAGCTGTGTGACCACCATGGCCAAGAA CAAGCCAACTCTTGATTTCGAACTGATCAAGACCGAGGCCAAGCA GCCTGCCACTCTGAGGAAGTACTGTATCGAAGCGAAGCTGACCAA CACCACTACCGAATCCCGCTGCCCGACCCAGGGCGAACCTTCCTTG AACGAAGAACAGGACAAGAGATTCGTGTGCAAGCATAGCATGGTC GACAGGGGATGGGGGAACGGATGTGGACTCTTTGGGAAGGGCGG AATCGTCACCTGTGCGATGTTCCGGTGCAAGAAGAACATGGAGGG GAAGGTCGTGCAGCCCGAAAATCTCGAGTACACTATCGTGATCAC CCCGCATTCCGGAGAGGAGCACGCCGTGGGCAACGACACCGGGA AGCACGGAAAGGAGATCAAAATTACCCCTCAATCCTCCACCACCG AAGCCGAATTGACTGGTTACGGTACCGTGACTATGGAGTGCTCGC CGCGGACTGGCTTGGACTTCAACGAGATGGTGCTGCTGCAAATGG AGAACAAGGCCTGGCTGGTGCACCGGCAGTGGTTTCTTGATCTGC CTCTGCCTTGGCTGCCCGGAGCCGACACCCAGGGTAGCAATTGGA TCCAGAAAGAGACACTCGTGACCTTTAAGAACCCGCACGCAAAGA AGCAGGATGTCGTGGTCCTGGGAAGCCAAGAAGGGGCAATGCATA CCGCACTCACTGGAGCCACTGAAATCCAGATGTCCTCCGGCAATCT GCTGTTCACCGGCCATCTGAAGTGCCGACTGCGCATGGACAAGCT CCAGCTTAAGGGAATGTCCTACTCCATGTGTACTGGAAAGTTCAA AGTCGTGAAGGAAATTGCCGAAACCCAGCACGGCACCATAGTGAT CCGGGTGCAGTACGAGGGCGACGGCTCACCCTGCAAAATCCCGTT CGAGATTATGGATCTCGAAAAGCGCCACGTGCTGGGCAGACTGAT TACCGTGAACCCTATCGTGACCGAGAAGGATTCCCCAGTGAACAT CGAGGCCGAACCGCCCTTCGGAGACTCGTATATCATCATCGGCGT GGAGCCCGGCCAGCTGAAGCTGAACTGGTTCAAGAAGGGGTCGAG CATCGGCCAGATGTTCGAGACTACCATGCGCGGCGCGAAGAGGAT GGCGATCCTGGGGGATACCGCTTGGGACTTCGGTTCCCTCGGCGG GGTGTTCACCTCGATTGGGAAGGCCCTCCACCAAGTGTTCGGTGCA ATCTACGGAGCGGCGTTCAGCGGAGTGTCGTGGACCATGAAGATT CTGATCGGCGTGATCATCACCTGGATTGGCATGAACTCCCGGTCTA CTAGCCTGTCGGTGACCCTGGTGCTGGTCGGAATCGTGACCTTGTA CCTGGGAGTGATGGTGCAAGCTTAG Dengue 2 prME ATGCTGAACATCCTGAACCGCAGAAGGAGAACCGCCGGTATTATT 233 (Jamaica/1409/1983) ATTATGATGATCCCCACCGTGATGGCATTCCACCTGACTACCCGCA ACGGAGAGCCGCATATGATCGTGGGCCGCCAGGAAAAGGGAAAG TCCCTGCTGTTCAAGACTGAGGACGGCGTGAACATGTGCACTCTCA TGGCCATCGACCTCGGCGAACTGTGCGAGGACACCATTACTTACA AGTGCCCGCTGCTGAGACAGAACGAGCCTGAGGACATCGACTGTT GGTGTAACTCGACCTCCACCTGGGTCACCTACGGAACGTGCGCCA CAACCGGAGAACACCGCCGGGAAAAGCGGAGCGTGGCTCTGGTG CCGCACGTCGGAATGGGTCTGGAGACTAGAACCGAAACCTGGATG TCATCCGAGGGGGCATGGAAACATGTGCAGCGAATCGAGACTTGG ATCCTGAGACACCCGGGCTTCACTATCATGGCGGCCATCCTTGCCT ACACCATTGGCACTACTCACTTCCAACGGGCGCTGATCTTCATACT GCTCACCGCGGTGGCCCCCTCCATGACGATGCGCTGCATCGGAAT CTCCAACCGGGACTTCGTGGAGGGCGTCAGCGGAGGCAGCTGGGT GGACATCGTGTTGGAGCACGGAAGCTGCGTGACCACCATGGCCAA GAACAAGCCCACTCTTGATTTTGAGCTGATCAAGACGGAAGCAAA GCAGCCGGCCACTCTGAGGAAGTACTGCATCGAGGCCAAGCTCAC CAACACAACCACCGAATCTCGGTGCCCGACCCAAGGAGAGCCATC ACTGAACGAGGAACAGGACAAGAGATTCCTGTGCAAACATTCGAT GGTGGACAGGGGATGGGGAAATGGTTGCGGCCTGTTCGGCAAAGG AGGCATTGTGACCTGTGCGATGTTCACTTGCAAGAAAAACATGGA GGGGAAGGTCGTGTTGCCGGAGAACCTGGAGTACACTATCGTGAT TACCCCGCACTCCGGGGAGGAACATGCCGTGGGAAATGACACCGG AAAGCACGGGAAGGAAATCAAAATCACGCCTCAGTCCTCAATCAC CGAAGCCGAGCTTACCGGCTACGGTACCGTGACCATGGAGTGCAG CCCTCGGACTGGACTGGACTTCAACGAGATGGTGCTGCTGCAAAT GGAAGATAAGGCCTGGCTGGTGCACCGGCAGTGGTTCTTGGATTT GCCACTGCCTTGGCTGCCCGGCGCGGATACCCAGGGTTCCAACTG GATTCAGAAGGAAACCCTCGTGACCTTCAAGAATCCTCACGCCAA GAAGCAGGACGTGGTGGTGCTGGGTTCCCAAGAAGGGGCCATGCA TACTGCCCTCACTGGAGCGACCGAAATCCAGATGTCGTCCGGCAA CCTCCTGTTCACCGGCCACCTGAAGTGCCGCCTGCGGATGGACAA GTTGCAGCTGAAGGGAATGAGCTACTCGATGTGTACCGGAAAGTT CAAGATCGTGAAGGAAATCGCCGAAACCCAGCACGGAACCATCGT CATTAGAGTGCAGTACGAAGGGGACGGCAGCCCGTGCAAGATCCC CTTCGAAATTATGGACCTGGAGAAGCGCCACGTGCTCGGAAGGCT CATCACTGTCAACCCAATCGTCACCGAAAAGGACTCCCCTGTGAA CATCGAAGCAGAGCCCCCTTTCGGGGACTCCTACATTATTATCGGC GTGGAGCCCGGCCAGCTGAAGCTGAACTGGTTCAAGAAGGGATCC TCGATCGGACAGATGTTCGAAACCACCATGCGGGGAGCCAAGCGG ATGGCTATTCTGGGAGATACCGCTTGGGATTTCGGCTCCCTCGGCG GCGTCTTTACTTCCATCGGGAAAGCGCTCCACCAAGTGTTTGGAGC CATCTACGGTGCCGCTTTTTCCGGGGTGTCATGGACCATGAAGATT CTTATCGGGGTCATTATTACTTGGATCGGCATGAACTCCCGGAGCA CCTCGCTGTCCGTGAGCCTCGTGCTCGTGGGGGTGGTCACTCTGTA TCTTGGTGCCATGGTGCAGGCCTAG Dengue 2 prME ATGCTTAACATCCTGAATAGAAGAAGAAGAACCGCCGGCATTATC 234 (Thailand/NGS- ATTATGATGATACCCACCGTGATGGCCTTCCACCTGACTACTCGCA C/1944) ACGGAGAGCCTCATATGATCGTGTCGCGGCAGGAGAAGGGAAAGT CCCTGCTGTTTAAGACGGAGGACGGCGTGAACATGTGCACTCTTAT GGCAATGGACCTTGGAGAGCTGTGCGAGGATACCATCACCTACAA GTGTCCGTTCCTGAAGCAAAACGAGCCTGAGGATATTGACTGCTG GTGCAACTCCACCTCAACCTGGGTCACATATGGGACCTGTACCACT ACTGGCGAACACCGCCGCGAGAAAAGAAGCGTGGCGTTGGTGCCT CACGTCGGCATGGGTCTGGAAACTCGGACCGAAACTTGGATGAGC TCAGAGGGGGCATGGAAGCACGCCCAGAGGATTGAAACCTGGATT CTGCGCCACCCTGGATTCACCATCATGGCGGCTATTCTGGCGTACA CTATTGGAACCACCCACTTTCAGCGGGCCCTTATCTTCATCCTCCT CACTGCCGTGGCGCCCTCCATGACTATGCGGTGTATCGGAATTTCC AACCGCGACTTCGTGGAAGGAGTGTCCGGAGGCTCCTGGGTCGAC ATTGTGCTGGAACATGGTTCATGCGTGACCACGATGGCCAAGAAC AAGCCCACCCTCGACTTCGAGCTGATCGAGACTGAAGCCAAGCAG CCGGCCACTCTGCGGAAGTACTGTATCGAGGCCAAGCTCACCAAC ACCACCACCGATTCCCGCTGCCCGACCCAAGGAGAACCTTCGCTC AACGAGGAGCAGGACAAGCGGTTCGTGTGCAAGCACAGCATGGTC GACAGGGGATGGGGGAATGGATGCGGTCTGTTCGGAAAGGGAGG CATTGTGACTTGTGCAATGTTCACTTGCAAGAAGAACATGAAGGG GAAGGTCGTGCAGCCGGAAAACCTGGAGTACACCATCGTGATCAC CCCTCATTCGGGCGAAGAACACGCTGTGGGGAATGATACCGGAAA GCACGGAAAGGAAATTAAGATCACACCCCAATCCAGCATCACTGA GGCAGAACTGACCGGCTACGGCACTGTGACCATGGAGTGCTCGCC TCGGACTGGCCTGGACTTCAACGAGATGGTGCTGCTCCAAATGGA AAACAAGGCCTGGCTGGTGCACAGACAGTGGTTCCTCGATTTGCC CTTGCCGTGGCTCCCTGGCGCCGACACCCAGGGATCTAACTGGATC CAGAAGGAAACCCTTGTGACCTTCAAGAACCCGCACGCTAAGAAA CAGGATGTGGTGGTGCTGGGAAGCCAGGAAGGAGCAATGCATACC GCGCTCACGGGTGCCACCGAGATCCAGATGAGCTCCGGGAACCTC CTGTTCACCGGTCACCTGAAGTGCCGACTCCGCATGGACAAACTG CAGCTCAAGGGGATGTCCTACTCCATGTGCACCGGGAAATTCAAG GTCGTGAAGGAGATCGCTGAGACTCAGCACGGTACTATCGTGATC CGGGTGCAGTATGAGGGAGATGGGAGCCCGTGCAAAATCCCATTT GAGATCATGGACTTGGAAAAGCGCCATGTGCTGGGTCGGCTGATT ACCGTGAACCCAATCGTCACCGAAAAGGACAGCCCCGTCAACATT GAAGCCGAACCACCCTTCGGAGACTCGTACATCATCATTGGCGTG GAACCGGGCCAGCTGAAGCTGAACTGGTTCAAAAAGGGGTCCTCT ATCGGCCAAATGATCGAAACCACCATGCGGGGAGCTAAGCGGATG GCGATTTTGGGAGACACTGCGTGGGACTTTGGCTCACTGGGGGGA GTGTTCACCAGCATCGGCAAAGCCCTGCACCAAGTGTTCGGTGCC ATCTACGGAGCCGCCTTCAGCGGAGTGTCCTGGATCATGAAGATC CTGATCGGCGTGATCATTACCTGGATCGGCATGAACTCCAGGTCCA CCTCGCTCTCCGTGTCGCTGGTGCTGGTCGGGGTCGTGACCCTGTA CCTGGGAGTGATGGTCCAGGCCTGA Dengue 2 prME ATGTTGAATATCCTGAACCGCCGCCGGAGAACTGCCGGAATTATC 235 (PuertoRico/PR159- ATTATGATGATCCCTACCGTGATGGCGTTCCACCTTACTACCCGGA S1/1969) ACGGGGAGCCTCACATGATCGTGTCACGCCAGGAGAAGGGGAAAT CCCTGCTGTTCAAGACCAAGGACGGTACCAACATGTGTACCCTGA TGGCGATGGACCTCGGAGAGCTGTGCGAGGACACCATCACCTACA AATGCCCGTTCCTGAAGCAGAACGAGCCGGAAGATATTGACTGTT GGTGCAACTCCACCTCCACTTGGGTCACCTACGGAACTTGCACCAC TACTGGGGAGCATAGACGGGAGAAGCGCTCCGTGGCCCTGGTGCC GCACGTCGGCATGGGACTGGAAACCAGAACCGAGACTTGGATGTC CAGCGAAGGCGCCTGGAAGCACGCCCAGCGGATTGAAACTTGGAT CCTGAGGCACCCGGGTTTTACCATTATGGCCGCTATCTTGGCATAC ACCATCGGCACCACCCACTTCCAACGCGTCCTGATCTTCATCCTGC TGACCGCCATTGCGCCCTCCATGACCATGCGGTGCATCGGAATCA GCAACCGCGACTTCGTGGAAGGCGTCAGCGGCGGTTCTTGGGTGG ACATCGTGTTGGAGCACGGATCGTGCGTGACCACCATGGCCAAGA ACAAGCCGACCCTCGATTTCGAGCTGATCAAGACTGAAGCCAAGC AGCCAGCTACCCTGCGGAAGTATTGCATCGAAGCCAAGCTCACTA ATACTACGACCGACAGCCGGTGTCCGACCCAAGGAGAGCCCACCC TGAATGAGGAACAAGACAAGCGCTTCGTGTGCAAGCATTCCATGG TGGACCGGGGCTGGGGAAACGGCTGCGGACTGTTCGGGAAAGGA GGAATTGTGACTTGCGCCATGTTCACTTGCAAGAAGAACATGGAG GGGAAGATCGTCCAGCCTGAGAACCTCGAGTACACGGTCGTGATT ACTCCGCACTCGGGAGAAGAACACGCCGTGGGCAACGACACCGG AAAGCATGGGAAGGAAGTGAAAATCACGCCCCAATCGTCGATTAC CGAGGCTGAGCTGACCGGCTACGGCACCGTGACCATGGAGTGCTC CCCGAGGACCGGACTGGACTTCAACGAAATGGTGCTGCTGCAGAT GAAGGACAAGGCCTGGCTGGTGCACCGCCAGTGGTTCCTCGACCT CCCACTCCCCTGGCTGCCCGGAGCGGATACGCAGGGATCCAACTG GATCCAGAAGGAAACTCTTGTGACCTTCAAGAACCCTCATGCCAA GAAGCAGGACGTGGTGGTCCTGGGATCCCAAGAGGGCGCGATGCA CACCGCACTGACCGGCGCCACCGAAATTCAGATGTCCTCCGGAAA CCTCCTGTTCACTGGCCACCTGAAGTGCAGACTCCGCATGGACAA GCTGCAGCTCAAGGGGATGAGCTACTCCATGTGTACCGGAAAATT CAAGGTCGTGAAGGAAATTGCAGAAACACAGCATGGGACAATTGT CATTCGGGTCCAGTACGAGGGCGATGGTTCACCGTGCAAGACTCC ATTCGAGATCATGGATCTGGAGAAAAGACACGTGCTGGGTCGGCT GACTACCGTGAACCCAATCGTGACTGAGAAGGACTCCCCCGTGAA CATCGAAGCCGAGCCTCCTTTTGGCGATTCCTACATCATCATTGGA GTGGAACCCGGACAGCTTAAGTTGGATTGGTTCAAGAAGGGCTCC TCGATCGGACAGATGTTCGAAACCACCATGCGCGGTGCCAAGCGA ATGGCCATCCTGGGGGACACCGCCTGGGACTTCGGTAGCCTGGGC GGAGTGTTTACCTCAATTGGAAAGGCTCTGCACCAAGTGTTTGGG GCGATCTACGGAGCGGCCTTCAGCGGTGTCTCCTGGACTATGAAG ATTCTCATCGGAGTGATAATCACCTGGATCGGCATGAACAGCCGG TCAACCAGCCTGTCCGTGTCCCTGGTGCTGGTCGGCATCGTGACTC TCTACCTCGGAGTGATGGTGCAGGCCTAG Dengue 2 prME ATGCTCAACATACTGAACAGACGGAGAAGGACCGCCGGTATTATT 236 (16681-PDK53) ATCATGATGATCCCTACTGTGATGGCATTCCACCTGACAACCCGCA ACGGAGAGCCCCACATGATCGTGTCACGCCAGGAGAAAGGGAAG TCACTGCTGTTCAAGACCGAAGTCGGCGTGAACATGTGTACCCTG ATGGCGATGGATCTTGGCGAACTGTGCGAGGACACCATCACGTAC AAGTGCCCCCTGTTGCGGCAAAACGAACCAGAGGACATCGACTGC TGGTGTAACTCCACCTCGACCTGGGTCACCTACGGAACCTGTACCA CTATGGGGGAACACCGGCGGGAGAAGCGCTCCGTGGCGCTCGTGC CTCATGTCGGCATGGGACTGGAGACTCGGACTGAAACCTGGATGT CGTCGGAGGGGGCCTGGAAGCACGTCCAGCGGATCGAGACTTGGA TCCTTCGCCATCCGGGCTTCACCATGATGGCCGCCATCCTGGCCTA CACCATCGGAACCACCCATTTCCAACGGGCCCTGATCCTGATCCTG TTGACTGCCGTGACCCCCTCCATGACTATGCGGTGCATTGGGATGT CGAACAGGGATTTCGTGGAGGGAGTCAGCGGTGGCAGCTGGGTGG ACATCGTGCTGGAACATGGATCCTGCGTGACTACCATGGCAAAGA ACAAGCCAACCCTCGATTTCGAACTGATCAAGACCGAGGCGAAAC AGCCGGCGACCCTGAGGAAGTACTGCATCGAGGCCAAGCTCACCA ACACCACTACCGAGAGCAGATGCCCTACCCAAGGGGAACCTTCCC TGAACGAGGAGCAGGACAAGAGATTCGTCTGCAAGCACTCCATGG TGGACCGCGGCTGGGGAAACGGATGCGGACTCTTCGGAAAGGGCG GTATTGTGACCTGTGCCATGTTCCGCTGCAAGAAAAACATGGAAG GGAAAGTGGTGCAGCCCGAGAACCTCGAGTACACTATCGTGATCA CACCGCACAGCGGAGAAGAACACGCCGTGGGCAACGACACTGGA AAGCACGGGAAGGAAATCAAGATCACCCCGCAATCCTCAATCACT GAGGCTGAGTTGACCGGCTACGGGACTATTACCATGGAATGCTCC CCACGAACGGGACTGGACTTCAACGAAATTGTGTTGCTCCAAATG GAAAACAAGGCCTGGCTCGTGCACCGGCAGTGGTTCCTGGATCTG CCCCTGCCGTGGCTGCCGGGTGCCGACACTCAGGGGAGCAACTGG ATTCAGAAGGAAACCCTTGTGACCTTCAAGAACCCCCACGCAAAG AAGCAGGACGTGGTGGTGCTGGGTAGCCAAGAAGGCGCCATGCAC ACGGCCCTGACCGGAGCGACCGAGATCCAGATGTCCAGCGGAAAT CTGCTCTTTACTGGTCATCTGAAGTGCAGACTTCGGATGGACAAGC TGCAACTGAAGGGAATGTCCTACTCAATGTGCACTGGAAAGTTCA AGGTCGTGAAGGAGATCGCCGAAACCCAGCACGGGACTATCGTCA TCCGCGTGCAGTACGAAGGAGATGGCTCCCCGTGCAAGATCCCTT TCGAAATCATGGACCTGGAGAAGCGCCACGTGTTGGGGCGCCTTA TTACTGTGAACCCCATCGTGACCGAGAAGGACTCCCCTGTCAACAT CGAGGCTGAACCGCCATTCGGAGATTCCTATATCATTATCGGAGTG GAACCGGGCCAGCTCAAGCTGAATTGGTTCAAGAAGGGATCCTCG ATTGGCCAGATGTTCGAAACGACTATGCGGGGCGCTAAGCGCATG GCCATCCTGGGCGATACTGCCTGGGATTTTGGTTCTCTGGGCGGAG TGTTCACCTCCATTGGAAAGGCCCTGCACCAAGTGTTCGGCGCCAT CTACGGTGCCGCGTTTAGCGGTGTCTCATGGACCATGAAAATCCTC ATTGGCGTGATCATTACCTGGATTGGCATGAACTCCAGAAGCACTT CCCTGTCCGTGACCCTGGTGCTCGTCGGAATTGTGACACTCTACCT CGGAGTGATGGTGCAGGCTTGA Dengue 2 prME ATGCTGAACATTTTGAACAGACGCCGAAGGACCGCAGGCATTATC 237 (Peru/IQT2913/1996) ATTATGATGATCCCTACCGTGATGGCCTTCCATCTGACTACTAGGA ACGGAGAGCCACATATGATCGTGTCGCGCCAGGAAAAGGGAAAG AGCCTGCTTTTTAAAACCAAGGACGGCACGAACATGTGCACCCTT ATGGCCATGGACCTGGGGGAGTTGTGCGAGGACACCATCACCTAC AAGTGCCCGTTCCTGAAGCAAAACGAGCCCGAAGATATTGACTGC TGGTGCAACTCCACCTCCACCTGGGTCACTTATGGGACTTGCACCA CCACCGGCGAACATCGCAGAGAAAAGAGAAGCGTGGCCCTGGTCC CCCACGTCGGGATGGGCCTCGAGACTCGGACCGAAACTTGGATGT CATCAGAGGGCGCATGGAAGCATGCTCAGCGGATCGAAACCTGGA TCCTGAGACACCCTGGTTTCACAATTATGGCCGCCATTCTTGCGTA CACGATCGGAACGACTCATTTCCAACGCGTGCTGATCTTCATTCTC CTGACCGCTATTGCGCCGTCCATGACTATGCGGTGCATCGGAATCT CAAACCGGGACTTCGTGGAAGGAGTGTCGGGAGGATCCTGGGTGG ACATTGTGCTGGAGCACGGTTCCTGCGTCACCACCATGGCCAAAA ACAAGCCTACCCTGGACTTCGAGCTGATCAAGACTGAGGCCAAGC AGCCCGCGACCCTCCGGAAGTACTGCATCGAGGCCAAGTTGACCA ACACTACTACCGATTCCCGGTGCCCGACCCAAGGAGAACCAACTC TGAACGAAGAACAGGATAAGCGGTTTGTGTGCAAGCACTCAATGG TGGACAGGGGATGGGGCAACGGCTGTGGACTGTTCGGAAAGGGTG GTATTGTGACCTGTGCAATGTTTACCTGTAAAAAGAATATGGAGG GGAAGATCGTGCAGCCTGAAAATCTCGAGTACACTGTCGTCATCA CCCCGCACTCGGGAGAGGAGCACGCTGTGGGCAACGACACCGGA AAGCACGGAAAGGAGGTCAAGATAACCCCGCAATCCTCCATTACG GAAGCCGAACTGACTGGTTACGGCACCGTGACTATGGAGTGCTCC CCTCGGACCGGCCTGGACTTCAACGAAATGGTGCTGCTCCAAATG GAAGATAAGGCCTGGCTGGTGCACAGGCAGTGGTTCCTGGATCTC CCGCTGCCGTGGCTGCCTGGCGCTGACACTCAGGGAAGCAACTGG ATCCAGAAGGAAACCCTCGTGACCTTTAAGAACCCCCACGCCAAG AAGCAGGATGTGGTGGTGTTGGGAAGCCAGGAGGGGGCCATGCAT ACTGCCCTCACCGGCGCGACCGAAATCCAGATGTCGTCCGGCAAT CTGCTGTTCACCGGACACCTCAAGTGTCGCCTTCGGATGGACAAGC TGCAGCTGAAGGGAATGAGCTACAGCATGTGCACCGGGAAGTTCA AGATCGTGAAGGAAATCGCCGAAACCCAGCACGGAACCATCGTGA TCCGGGTGCAGTACGAGGGCGACGGTTCTCCCTGCAAAATCCCCTT CGAAATCATGGATCTGGAGAAGAGACACGTCCTGGGTCGCCTGAT CACCGTGAACCCCATTGTGACTGAGAAGGACTCCCCAGTGAACAT CGAAGCGGAGCCCCCATTCGGAGACAGCTACATTATCATTGGTGC CGAACCGGGGCAGCTGAAACTGGACTGGTTCAAGAAGGGCAGCTC GATTGGCCAAATGTTCGAAACGACAATGCGGGGCGCAAAGCGCAT GGCCATCCTGGGAGACACTGCCTGGGACTTCGGGTCCCTTGGGGG GGTGTTCACCTCGATCGGAAAAGCCTTGCACCAAGTGTTCGGCGC AATCTACGGCGCCGCGTTCTCGGGAGTCTCCTGGACTATGAAGATC CTGATCGGTGTCATCATCACCTGGATCGGGATGAACTCCCGGTCCA CTTCCCTCTCGGTGTCACTCGTGCTTGTGGGAATTGTCACCCTGTA CCTCGGAGTGATGGTGCAGGCCTGA Dengue 2 prME ATGCTGAATATTCTGAACCGACGCCGCCGCACTGCCGGAATCATT 238 (Thailand/PUO- ATCATGATGATCCCTACCGTGATGGCGTTCCATCTCACCACTCGGA 218/1980) ATGGCGAACCCCATATGATCGTGTCGAGACAGGAAAAGGGAAAG AGCCTTTTGTTCAAAACTGAAGATGGAGTGAACATGTGCACTCTCA TGGCAATGGATCTGGGCGAACTGTGCGAAGATACCATCACTTACA AGTGTCCGCTGTTGAGACAGAACGAGCCTGAGGACATCGACTGCT GGTGTAACAGCACTTCCACCTGGGTCACCTACGGCACTTGCACTAC CACCGGAGAACACCGGCGCGAGAAGAGGAGCGTGGCTCTTGTGCC GCACGTCGGCATGGGACTCGAGACTCGGACCGAAACCTGGATGTC ATCCGAAGGAGCCTGGAAACACGCCCAACGGATCGAAATTTGGAT CCTGAGACACCCCGGTTTCACTATCATGGCCGCAATCCTGGCGTAC ACTATTGGCACCACGCACTTCCAGAGGGCCCTCATTTTCATCCTCC TGACTGCCGTGGCGCCATCCATGACCATGAGATGTATTGGCATTTC CAACCGCGATTTCGTGGAGGGAGTGTCCGGAGGATCCTGGGTCGA CATCGTGCTGGAACACGGATCTTGCGTCACCACCATGGCTAAGAA CAAGCCCACCCTCGACTTCGAGCTGATCAAGACAGAAGCCAAGCA GCCGGCCACCCTCCGCAAGTATTGCATTGAAGCCAAGCTTACCAA CACCACCACCGAGTCGCGGTGCCCAACCCAAGGAGAGCCGAGCCT CAATGAGGAACAGGACAAGCGCTTCGTGTGCAAACACAGCATGGT CGACCGGGGTTGGGGCAACGGATGTGGCCTGTTCGGGAAGGGTGG CATTGTGACTTGCGCAATGTTCACTTGCAAGAAGAACATGGAGGG GAAAGTGGTGCAACCCGAGAACCTGGAGTACACCATCGTCGTGAC CCCACACTCCGGAGAGGAGCACGCCGTGGGAAACGACACGGGGA AGCATGGAAAGGAGATCAAGGTCACACCCCAATCATCTATTACCG AGGCCGAACTGACCGGATACGGTACTGTGACGATGGAGTGCAGCC CGAGGACTGGACTGGACTTCAACGAAATGGTGCTGCTGCAAATGG AGAACAAGGCCTGGCTCGTGCACCGGCAGTGGTTTCTGGATCTCC CACTGCCGTGGTTGCCGGGAGCCGACACCCAGGGGTCGAACTGGA TCCAGAAGGAAACTCTTGTGACGTTTAAGAATCCTCACGCGAAGA AGCAGGACGTGGTGGTCCTGGGATCGCAGGAAGGAGCTATGCACA CCGCTCTGACCGGCGCCACTGAGATCCAGATGTCCTCGGGCAACC TCCTGTTCACCGGTCATCTGAAGTGCCGGCTGCGGATGGACAAATT GCAGCTGAAGGGGATGTCCTACTCCATGTGCACCGGGAAGTTCAA GGTCGTGAAGGAGATCGCGGAAACTCAGCACGGCACCATTGTCAT TAGAGTGCAGTACGAGGGAGATGGTTCACCGTGCAAGATACCGTT CGAAATCATGGACCTGGAAAAGAGACATGTCTTGGGACGCCTGAT CACTGTGAACCCTATCGTGACCGAAAAGGACTCCCCTGTGAACAT CGAGGCGGAGCCGCCTTTCGGCGACTCCTACATCATTATCGGAGT GGAGCCCGGGCAGCTGAAGCTCAACTGGTTTAAGAAGGGGTCCAG CATCGGCCAGATGTTCGAAACCACCATGCGGGGGGCGAAGAGGAT GGCGATCCTGGGAGACACCGCCTGGGATTTCGGTTCACTGGGCGG AGTGTTCACCTCCATCGGAAAGGCCCTGCACCAAGTGTTCGGCGC AATCTACGGTGCTGCCTTCTCGGGAGTCTCCTGGACCATGAAGATC CTGATCGGCGTGATTATCACATGGATCGGCATGAACAGCCGGTCA ACCTCCCTTTCCGTGTCCCTGGTGCTGGTCGGCATCGTGACTCTGT ACCTGGGCGTGATGGTGCAGGCCTGA Dengue 2 prME ATGCTGAACATTCTGAACCGGAGAAGAAGAACCGCCGGCATTATT 239 (D2Y98P) with ATCATGATGATTCCCACTGTGATGGCATTTCACCTGACCACCCGGA native leader ACGGAGAACCTCATATGATCGTGTCGAGACAGGAGAAGGGAAAG TCCCTGCTGTTCAAGACAGAAAACGGAGTGAACATGTGCACCCTG ATGGCCATGGATCTCGGCGAACTGTGCGAGGATACTATCACCTAC AACTGTCCGTTGCTGCGCCAAAACGAGCCGGAGGACATCGACTGC TGGTGTAACTCCACGTCGACCTGGGTCACCTACGGCACTTGCACCG CGACCGGCGAACACAGAAGAGAGAAACGCTCCGTCGCTCTGGTGC CGCACGTCGGGATGGGGCTTGAAACCCGGACTGAAACCTGGATGA GCTCGGAGGGCGCTTGGAAGCATGCCCAGCGCATCGAAACTTGGG TGCTGAGGCATCCAGGCTTCACAATCATGGCCGCCATCCTCGCGTA CACCATCGGTACTACGTACTTCCAGCGGGTGTTGATCTTCATTCTG CTGACCGCCGTGGCCCCTAGCATGACCATGCGGTGCATCGGGATC TCCAACCGCGATTTCGTGGAGGGGGTGTCCGGTGGAAGCTGGGTG GACATTGTGCTGGAGCACGGCTCGTGCGTGACCACCATGGCCAAG AACAAGCCCACCCTTGATTTTGAGCTGATCAAGACCGAAGCGAAA CACCCCGCGACCCTCCGGAAGTACTGCATTGAAGCCAAGCTCACC AACACTACCACGGCCTCCCGGTGCCCTACCCAAGGAGAACCTTCC TTGAACGAAGAACAGGACAAGCGCTTCGTGTGCAAGCATTCAATG GTGGACCGGGGCTGGGGAAATGGCTGTGGCCTCTTCGGAAAAGGC GGAATTGTGACTTGCGCAATGTTCACTTGCAAGAAGAACATGGAG GGAAAGATTGTGCAGCCCGAGAACCTCGAGTACACTATTGTCATC ACTCCCCACTCCGGCGAAGAAAACGCTGTCGGCAACGACACCGGA AAGCATGGAAAGGAGATCAAGGTCACCCCGCAATCCTCAATTACT GAGGCAGAACTGACCGGTTACGGAACTGTGACTATGGAGTGTTCC CCTCGCACCGGCCTCGATTTCAACGAGATGGTGCTGCTGCAAATG GAGAACAAGGCCTGGCTGGTGCACCGGCAGTGGTTCCTCGATTTG CCCCTGCCGTGGCTGCCGGGAGCCGACACTCAGGGATCCAACTGG ATCCAGAAAGAAACCCTCGTGACCTTCAAAAACCCCCACGCGAAG AAGCAGGACGTGGTGGTGCTGGGTTCCCAAGAAGGGGCGATGCAT ACCGCCCTGACTGGTGCTACCGAAATCCAGATGTCAAGCGGAAAT CTCCTGTTTACCGGTCACCTGAAGTGCAGGCTCCGGATGGACAAGT TGCAGCTGAAGGGGATGTCGTACAGCATGTGTACTGGGAAGTTCA AGGTCGTGAAGGAGATTGCCGAAACCCAGCACGGAACCATAGTCA TCAGGGTCCAGTACGAGGGCGACGGCAGCCCTTGCAAGATCCCGT TCGAGATCATGGATCTGGAGAAGCGACACGTGCTGGGCCGGCTTA TCACTGTGAATCCAATCGTGACCGAGAAAGACTCGCCCGTGAACA TCGAAGCCGAGCCGCCGTTCGGCGACTCATACATCATCATCGGCG TGGAACCAGGACAGCTGAAGCTGTCATGGTTCAAGAAGGGTTCCA GCATTGGTCAGATGTTCGAAACAACGATGCGCGGAGCCAAGCGCA TGGCTATCCTTGGGGACACCGCCTGGGACTTCGGGTCGCTGGGAG GAGTGTTTACCAGCATCGGAAAGGCCCTGCACCAAGTGTTCGGTG CCATCTACGGAGCCGCATTTTCCGGAGTGTCGTGGACTATGAAGAT TCTGATCGGCGTCGTGATTACCTGGATCGGGATGAACTCCAGGTCT ACTTCCCTCTCCGTGAGCCTGGTGCTGGTCGGCGTGGTCACCCTGT ATCTGGGCGTGATGGTCCAGGCTTAG mRNA Sequences DEN-1 AGUUGUUAGUCUACGUGGACCGACAAGAACAGUUUCGAAUCGG 240 (NC_001477.1) AAGCUUGCUUAACGUAGUUCUAACAGUUUUUUAUUAGAGAGCA GAUCUCUGAUGAACAACCAACGGAAAAAGACGGGUCGACCGUCU UUCAAUAUGCUGAAACGCGCGAGAAACCGCGUGUCAACUGUUUC ACAGUUGGCGAAGAGAUUCUCAAAAGGAUUGCUUUCAGGCCAA GGACCCAUGAAAUUGGUGAUGGCUUUUAUAGCAUUCCUAAGAU UUCUAGCCAUACCUCCAACAGCAGGAAUUUUGGCUAGAUGGGGC UCAUUCAAGAAGAAUGGAGCGAUCAAAGUGUUACGGGGUUUCA AGAAAGAAAUCUCAAACAUGUUGAACAUAAUGAACAGGAGGAA AAGAUCUGUGACCAUGCUCCUCAUGCUGCUGCCCACAGCCCUGG CGUUCCAUCUGACCACCCGAGGGGGAGAGCCGCACAUGAUAGUU AGCAAGCAGGAAAGAGGAAAAUCACUUUUGUUUAAGACCUCUG CAGGUGUCAACAUGUGCACCCUUAUUGCAAUGGAUUUGGGAGA GUUAUGUGAGGACACAAUGACCUACAAAUGCCCCCGGAUCACUG AGACGGAACCAGAUGACGUUGACUGUUGGUGCAAUGCCACGGAG ACAUGGGUGACCUAUGGAACAUGUUCUCAAACUGGUGAACACCG ACGAGACAAACGUUCCGUCGCACUGGCACCACACGUAGGGCUUG GUCUAGAAACAAGAACCGAAACGUGGAUGUCCUCUGAAGGCGCU UGGAAACAAAUACAAAAAGUGGAGACCUGGGCUCUGAGACACCC AGGAUUCACGGUGAUAGCCCUUUUUCUAGCACAUGCCAUAGGAA CAUCCAUCACCCAGAAAGGGAUCAUUUUUAUUUUGCUGAUGCUG GUAACUCCAUCCAUGGCCAUGCGGUGCGUGGGAAUAGGCAACAG AGACUUCGUGGAAGGACUGUCAGGAGCUACGUGGGUGGAUGUG GUACUGGAGCAUGGAAGUUGCGUCACUACCAUGGCAAAAGACAA ACCAACACUGGACAUUGAACUCUUGAAGACGGAGGUCACAAACC CUGCCGUCCUGCGCAAACUGUGCAUUGAAGCUAAAAUAUCAAAC ACCACCACCGAUUCGAGAUGUCCAACACAAGGAGAAGCCACGCU GGUGGAAGAACAGGACACGAACUUUGUGUGUCGACGAACGUUC GUGGACAGAGGCUGGGGCAAUGGUUGUGGGCUAUUCGGAAAAG GUAGCUUAAUAACGUGUGCUAAGUUUAAGUGUGUGACAAAACU GGAAGGAAAGAUAGUCCAAUAUGAAAACUUAAAAUAUUCAGUG AUAGUCACCGUACACACUGGAGACCAGCACCAAGUUGGAAAUGA GACCACAGAACAUGGAACAACUGCAACCAUAACACCUCAAGCUC CCACGUCGGAAAUACAGCUGACAGACUACGGAGCUCUAACAUUG GAUUGUUCACCUAGAACAGGGCUAGACUUUAAUGAGAUGGUGU UGUUGACAAUGAAAAAAAAAUCAUGGCUCGUCCACAAACAAUG GUUUCUAGACUUACCACUGCCUUGGACCUCGGGGGCUUCAACAU CCCAAGAGACUUGGAAUAGACAAGACUUGCUGGUCACAUUUAAG ACAGCUCAUGCAAAAAAGCAGGAAGUAGUCGUACUAGGAUCACA AGAAGGAGCAAUGCACACUGCGUUGACUGGAGCGACAGAAAUCC AAACGUCUGGAACGACAACAAUUUUUGCAGGACACCUGAAAUGC AGAUUAAAAAUGGAUAAACUGAUUUUAAAAGGGAUGUCAUAUG UAAUGUGCACAGGGUCAUUCAAGUUAGAGAAGGAAGUGGCUGA GACCCAGCAUGGAACUGUUCUAGUGCAGGUUAAAUACGAAGGA ACAGAUGCACCAUGCAAGAUCCCCUUCUCGUCCCAAGAUGAGAA GGGAGUAACCCAGAAUGGGAGAUUGAUAACAGCCAACCCCAUAG UCACUGACAAAGAAAAACCAGUCAACAUUGAAGCGGAGCCACCU UUUGGUGAGAGCUACAUUGUGGUAGGAGCAGGUGAAAAAGCUU UGAAACUAAGCUGGUUCAAGAAGGGAAGCAGUAUAGGGAAAAU GUUUGAAGCAACUGCCCGUGGAGCACGAAGGAUGGCCAUCCUGG GAGACACUGCAUGGGACUUCGGUUCUAUAGGAGGGGUGUUCAC GUCUGUGGGAAAACUGAUACACCAGAUUUUUGGGACUGCGUAU GGAGUUUUGUUCAGCGGUGUUUCUUGGACCAUGAAGAUAGGAA UAGGGAUUCUGCUGACAUGGCUAGGAUUAAACUCAAGGAGCAC GUCCCUUUCAAUGACGUGUAUCGCAGUUGGCAUGGUCACACUGU ACCUAGGAGUCAUGGUUCAGGCGGACUCGGGAUGUGUAAUCAAC UGGAAAGGCAGAGAACUCAAAUGUGGAAGCGGCAUUUUUGUCA CCAAUGAAGUCCACACCUGGACAGAGCAAUAUAAAUUCCAGGCC GACUCCCCUAAGAGACUAUCAGCGGCCAUUGGGAAGGCAUGGGA GGAGGGUGUGUGUGGAAUUCGAUCAGCCACUCGUCUCGAGAACA UCAUGUGGAAGCAAAUAUCAAAUGAAUUAAACCACAUCUUACU UGAAAAUGACAUGAAAUUUACAGUGGUCGUAGGAGACGUUAGU GGAAUCUUGGCCCAAGGAAAGAAAAUGAUUAGGCCACAACCCAU GGAACACAAAUACUCGUGGAAAAGCUGGGGAAAAGCCAAAAUC AUAGGAGCAGAUGUACAGAAUACCACCUUCAUCAUCGACGGCCC AAACACCCCAGAAUGCCCUGAUAACCAAAGAGCAUGGAACAUUU GGGAAGUUGAAGACUAUGGAUUUGGAAUUUUCACGACAAACAU AUGGUUGAAAUUGCGUGACUCCUACACUCAAGUGUGUGACCACC GGCUAAUGUCAGCUGCCAUCAAGGAUAGCAAAGCAGUCCAUGCU GACAUGGGGUACUGGAUAGAAAGUGAAAAGAACGAGACUUGGA AGUUGGCAAGAGCCUCCUUCAUAGAAGUUAAGACAUGCAUCUGG CCAAAAUCCCACACUCUAUGGAGCAAUGGAGUCCUGGAAAGUGA GAUGAUAAUCCCAAAGAUAUAUGGAGGACCAAUAUCUCAGCACA ACUACAGACCAGGAUAUUUCACACAAACAGCAGGGCCGUGGCAC UUGGGCAAGUUAGAACUAGAUUUUGAUUUAUGUGAAGGUACCA CUGUUGUUGUGGAUGAACAUUGUGGAAAUCGAGGACCAUCUCU UAGAACCACAACAGUCACAGGAAAGACAAUCCAUGAAUGGUGCU GUAGAUCUUGCACGUUACCCCCCCUACGUUUCAAAGGAGAAGAC GGGUGCUGGUACGGCAUGGAAAUCAGACCAGUCAAGGAGAAGG AAGAGAACCUAGUUAAGUCAAUGGUCUCUGCAGGGUCAGGAGA AGUGGACAGUUUUUCACUAGGACUGCUAUGCAUAUCAAUAAUG AUCGAAGAGGUAAUGAGAUCCAGAUGGAGCAGAAAAAUGCUGA UGACUGGAACAUUGGCUGUGUUCCUCCUUCUCACAAUGGGACAA UUGACAUGGAAUGAUCUGAUCAGGCUAUGUAUCAUGGUUGGAG CCAACGCUUCAGACAAGAUGGGGAUGGGAACAACGUACCUAGCU UUGAUGGCCACUUUCAGAAUGAGACCAAUGUUCGCAGUCGGGCU ACUGUUUCGCAGAUUAACAUCUAGAGAAGUUCUUCUUCUUACAG UUGGAUUGAGUCUGGUGGCAUCUGUAGAACUACCAAAUUCCUU AGAGGAGCUAGGGGAUGGACUUGCAAUGGGCAUCAUGAUGUUG AAAUUACUGACUGAUUUUCAGUCACAUCAGCUAUGGGCUACCUU GCUGUCUUUAACAUUUGUCAAAACAACUUUUUCAUUGCACUAUG CAUGGAAGACAAUGGCUAUGAUACUGUCAAUUGUAUCUCUCUUC CCUUUAUGCCUGUCCACGACUUCUCAAAAAACAACAUGGCUUCC GGUGUUGCUGGGAUCUCUUGGAUGCAAACCACUAACCAUGUUUC UUAUAACAGAAAACAAAAUCUGGGGAAGGAAAAGCUGGCCUCU CAAUGAAGGAAUUAUGGCUGUUGGAAUAGUUAGCAUUCUUCUA AGUUCACUUCUCAAGAAUGAUGUGCCACUAGCUGGCCCACUAAU AGCUGGAGGCAUGCUAAUAGCAUGUUAUGUCAUAUCUGGAAGC UCGGCCGAUUUAUCACUGGAGAAAGCGGCUGAGGUCUCCUGGGA AGAAGAAGCAGAACACUCUGGUGCCUCACACAACAUACUAGUGG AGGUCCAAGAUGAUGGAACCAUGAAGAUAAAGGAUGAAGAGAG AGAUGACACACUCACCAUUCUCCUCAAAGCAACUCUGCUAGCAA UCUCAGGGGUAUACCCAAUGUCAAUACCGGCGACCCUCUUUGUG UGGUAUUUUUGGCAGAAAAAGAAACAGAGAUCAGGAGUGCUAU GGGACACACCCAGCCCUCCAGAAGUGGAAAGAGCAGUCCUUGAU GAUGGCAUUUAUAGAAUUCUCCAAAGAGGAUUGUUGGGCAGGU CUCAAGUAGGAGUAGGAGUUUUUCAAGAAGGCGUGUUCCACAC AAUGUGGCACGUCACCAGGGGAGCUGUCCUCAUGUACCAAGGGA AGAGACUGGAACCAAGUUGGGCCAGUGUCAAAAAAGACUUGAU CUCAUAUGGAGGAGGUUGGAGGUUUCAAGGAUCCUGGAACGCG GGAGAAGAAGUGCAGGUGAUUGCUGUUGAACCGGGGAAGAACC CCAAAAAUGUACAGACAGCGCCGGGUACCUUCAAGACCCCUGAA GGCGAAGUUGGAGCCAUAGCUCUAGACUUUAAACCCGGCACAUC UGGAUCUCCUAUCGUGAACAGAGAGGGAAAAAUAGUAGGUCUU UAUGGAAAUGGAGUGGUGACAACAAGUGGUACCUACGUCAGUG CCAUAGCUCAAGCUAAAGCAUCACAAGAAGGGCCUCUACCAGAG AUUGAGGACGAGGUGUUUAGGAAAAGAAACUUAACAAUAAUGG ACCUACAUCCAGGAUCGGGAAAAACAAGAAGAUACCUUCCAGCC AUAGUCCGUGAGGCCAUAAAAAGAAAGCUGCGCACGCUAGUCUU AGCUCCCACAAGAGUUGUCGCUUCUGAAAUGGCAGAGGCGCUCA AGGGAAUGCCAAUAAGGUAUCAGACAACAGCAGUGAAGAGUGA ACACACGGGAAAGGAGAUAGUUGACCUUAUGUGUCACGCCACUU UCACUAUGCGUCUCCUGUCUCCUGUGAGAGUUCCCAAUUAUAAU AUGAUUAUCAUGGAUGAAGCACAUUUUACCGAUCCAGCCAGCAU AGCAGCCAGAGGGUAUAUCUCAACCCGAGUGGGUAUGGGUGAA GCAGCUGCGAUUUUCAUGACAGCCACUCCCCCCGGAUCGGUGGA GGCCUUUCCACAGAGCAAUGCAGUUAUCCAAGAUGAGGAAAGAG ACAUUCCUGAAAGAUCAUGGAACUCAGGCUAUGACUGGAUCACU GAUUUCCCAGGUAAAACAGUCUGGUUUGUUCCAAGCAUCAAAUC AGGAAAUGACAUUGCCAACUGUUUAAGAAAGAAUGGGAAACGG GUGGUCCAAUUGAGCAGAAAAACUUUUGACACUGAGUACCAGA AAACAAAAAAUAACGACUGGGACUAUGUUGUCACAACAGACAU AUCCGAAAUGGGAGCAAACUUCCGAGCCGACAGGGUAAUAGACC CGAGGCGGUGCCUGAAACCGGUAAUACUAAAAGAUGGCCCAGAG CGUGUCAUUCUAGCCGGACCGAUGCCAGUGACUGUGGCUAGCGC CGCCCAGAGGAGAGGAAGAAUUGGAAGGAACCAAAAUAAGGAA GGCGAUCAGUAUAUUUACAUGGGACAGCCUCUAAACAAUGAUG AGGACCACGCCCAUUGGACAGAAGCAAAAAUGCUCCUUGACAAC AUAAACACACCAGAAGGGAUUAUCCCAGCCCUCUUUGAGCCGGA GAGAGAAAAGAGUGCAGCAAUAGACGGGGAAUACAGACUACGG GGUGAAGCGAGGAAAACGUUCGUGGAGCUCAUGAGAAGAGGAG AUCUACCUGUCUGGCUAUCCUACAAAGUUGCCUCAGAAGGCUUC CAGUACUCCGACAGAAGGUGGUGCUUUGAUGGGGAAAGGAACA ACCAGGUGUUGGAGGAGAACAUGGACGUGGAGAUCUGGACAAA AGAAGGAGAAAGAAAGAAACUACGACCCCGCUGGCUGGAUGCCA GAACAUACUCUGACCCACUGGCUCUGCGCGAAUUCAAAGAGUUC GCAGCAGGAAGAAGAAGCGUCUCAGGUGACCUAAUAUUAGAAA UAGGGAAACUUCCACAACAUUUAACGCAAAGGGCCCAGAACGCC UUGGACAAUCUGGUUAUGUUGCACAACUCUGAACAAGGAGGAA AAGCCUAUAGACACGCCAUGGAAGAACUACCAGACACCAUAGAA ACGUUAAUGCUCCUAGCUUUGAUAGCUGUGCUGACUGGUGGAG UGACGUUGUUCUUCCUAUCAGGAAGGGGUCUAGGAAAAACAUCC AUUGGCCUACUCUGCGUGAUUGCCUCAAGUGCACUGUUAUGGAU GGCCAGUGUGGAACCCCAUUGGAUAGCGGCCUCUAUCAUACUGG AGUUCUUUCUGAUGGUGUUGCUUAUUCCAGAGCCGGACAGACAG CGCACUCCACAAGACAACCAGCUAGCAUACGUGGUGAUAGGUCU GUUAUUCAUGAUAUUGACAGUGGCAGCCAAUGAGAUGGGAUUA CUGGAAACCACAAAGAAGGACCUGGGGAUUGGUCAUGCAGCUGC UGAAAACCACCAUCAUGCUGCAAUGCUGGACGUAGACCUACAUC CAGCUUCAGCCUGGACUCUCUAUGCAGUGGCCACAACAAUUAUC ACUCCCAUGAUGAGACACACAAUUGAAAACACAACGGCAAAUAU UUCCCUGACAGCUAUUGCAAACCAGGCAGCUAUAUUGAUGGGAC UUGACAAGGGAUGGCCAAUAUCAAAGAUGGACAUAGGAGUUCC ACUUCUCGCCUUGGGGUGCUAUUCUCAGGUGAACCCGCUGACGC UGACAGCGGCGGUAUUGAUGCUAGUGGCUCAUUAUGCCAUAAU UGGACCCGGACUGCAAGCAAAAGCUACUAGAGAAGCUCAAAAAA GGACAGCAGCCGGAAUAAUGAAAAACCCAACUGUCGACGGGAUC GUUGCAAUAGAUUUGGACCCUGUGGUUUACGAUGCAAAAUUUG AAAAACAGCUAGGCCAAAUAAUGUUGUUGAUACUUUGCACAUC ACAGAUCCUCCUGAUGCGGACCACAUGGGCCUUGUGUGAAUCCA UCACACUAGCCACUGGACCUCUGACUACGCUUUGGGAGGGAUCU CCAGGAAAAUUCUGGAACACCACGAUAGCGGUGUCCAUGGCAAA CAUUUUUAGGGGAAGUUAUCUAGCAGGAGCAGGUCUGGCCUUU UCAUUAAUGAAAUCUCUAGGAGGAGGUAGGAGAGGCACGGGAG CCCAAGGGGAAACACUGGGAGAAAAAUGGAAAAGACAGCUAAA CCAAUUGAGCAAGUCAGAAUUCAACACUUACAAAAGGAGUGGG AUUAUAGAGGUGGAUAGAUCUGAAGCCAAAGAGGGGUUAAAAA GAGGAGAAACGACUAAACACGCAGUGUCGAGAGGAACGGCCAAA CUGAGGUGGUUUGUGGAGAGGAACCUUGUGAAACCAGAAGGGA AAGUCAUAGACCUCGGUUGUGGAAGAGGUGGCUGGUCAUAUUA UUGCGCUGGGCUGAAGAAAGUCACAGAAGUGAAAGGAUACACG AAAGGAGGACCUGGACAUGAGGAACCAAUCCCAAUGGCAACCUA UGGAUGGAACCUAGUAAAGCUAUACUCCGGGAAAGAUGUAUUC UUUACACCACCUGAGAAAUGUGACACCCUCUUGUGUGAUAUUGG UGAGUCCUCUCCGAACCCAACUAUAGAAGAAGGAAGAACGUUAC GUGUUCUAAAGAUGGUGGAACCAUGGCUCAGAGGAAACCAAUU UUGCAUAAAAAUUCUAAAUCCCUAUAUGCCGAGUGUGGUAGAA ACUUUGGAGCAAAUGCAAAGAAAACAUGGAGGAAUGCUAGUGC GAAAUCCACUCUCAAGAAACUCCACUCAUGAAAUGUACUGGGUU UCAUGUGGAACAGGAAACAUUGUGUCAGCAGUAAACAUGACAU CUAGAAUGCUGCUAAAUCGAUUCACAAUGGCUCACAGGAAGCCA ACAUAUGAAAGAGACGUGGACUUAGGCGCUGGAACAAGACAUG UGGCAGUAGAACCAGAGGUGGCCAACCUAGAUAUCAUUGGCCAG AGGAUAGAGAAUAUAAAAAAUGAACACAAAUCAACAUGGCAUU AUGAUGAGGACAAUCCAUACAAAACAUGGGCCUAUCAUGGAUCA UAUGAGGUCAAGCCAUCAGGAUCAGCCUCAUCCAUGGUCAAUGG UGUGGUGAGACUGCUAACCAAACCAUGGGAUGUCAUUCCCAUGG UCACACAAAUAGCCAUGACUGACACCACACCCUUUGGACAACAG AGGGUGUUUAAAGAGAAAGUUGACACGCGUACACCAAAAGCGA AACGAGGCACAGCACAAAUUAUGGAGGUGACAGCCAGGUGGUU AUGGGGUUUUCUCUCUAGAAACAAAAAACCCAGAAUCUGCACAA GAGAGGAGUUCACAAGAAAAGUCAGGUCAAACGCAGCUAUUGG AGCAGUGUUCGUUGAUGAAAAUCAAUGGAACUCAGCAAAAGAG GCAGUGGAAGAUGAACGGUUCUGGGACCUUGUGCACAGAGAGA GGGAGCUUCAUAAACAAGGAAAAUGUGCCACGUGUGUCUACAAC AUGAUGGGAAAGAGAGAGAAAAAAUUAGGAGAGUUCGGAAAGG CAAAAGGAAGUCGCGCAAUAUGGUACAUGUGGUUGGGAGCGCG CUUUUUAGAGUUUGAAGCCCUUGGUUUCAUGAAUGAAGAUCAC UGGUUCAGCAGAGAGAAUUCACUCAGUGGAGUGGAAGGAGAAG GACUCCACAAACUUGGAUACAUACUCAGAGACAUAUCAAAGAUU CCAGGGGGAAAUAUGUAUGCAGAUGACACAGCCGGAUGGGACAC AAGAAUAACAGAGGAUGAUCUUCAGAAUGAGGCCAAAAUCACU GACAUCAUGGAACCUGAACAUGCCCUAUUGGCCACGUCAAUCUU UAAGCUAACCUACCAAAACAAGGUAGUAAGGGUGCAGAGACCAG CGAAAAAUGGAACCGUGAUGGAUGUCAUAUCCAGACGUGACCAG AGAGGAAGUGGACAGGUUGGAACCUAUGGCUUAAACACCUUCAC CAACAUGGAGGCCCAACUAAUAAGACAAAUGGAGUCUGAGGGA AUCUUUUCACCCAGCGAAUUGGAAACCCCAAAUCUAGCCGAAAG AGUCCUCGACUGGUUGAAAAAACAUGGCACCGAGAGGCUGAAAA GAAUGGCAAUCAGUGGAGAUGACUGUGUGGUGAAACCAAUCGA UGACAGAUUUGCAACAGCCUUAACAGCUUUGAAUGACAUGGGA AAGGUAAGAAAAGACAUACCGCAAUGGGAACCUUCAAAAGGAU GGAAUGAUUGGCAACAAGUGCCUUUCUGUUCACACCAUUUCCAC CAGCUGAUUAUGAAGGAUGGGAGGGAGAUAGUGGUGCCAUGCC GCAACCAAGAUGAACUUGUAGGUAGGGCCAGAGUAUCACAAGGC GCCGGAUGGAGCUUGAGAGAAACUGCAUGCCUAGGCAAGUCAUA UGCACAAAUGUGGCAGCUGAUGUACUUCCACAGGAGAGACUUGA GAUUAGCGGCUAAUGCUAUCUGUUCAGCCGUUCCAGUUGAUUGG GUCCCAACCAGCCGCACCACCUGGUCGAUCCAUGCCCACCAUCA AUGGAUGACAACAGAAGACAUGUUGUCAGUGUGGAAUAGGGUU UGGAUAGAGGAAAACCCAUGGAUGGAGGACAAGACUCAUGUGU CCAGUUGGGAAGACGUUCCAUACCUAGGAAAAAGGGAAGAUCA AUGGUGUGGUUCCCUAAUAGGCUUAACAGCACGAGCCACCUGGG CCACCAACAUACAAGUGGCCAUAAACCAAGUGAGAAGGCUCAUU GGGAAUGAGAAUUAUCUAGACUUCAUGACAUCAAUGAAGAGAU UCAAAAACGAGAGUGAUCCCGAAGGGGCACUCUGGUAAGCCAAC UCAUUCACAAAAUAAAGGAAAAUAAAAAAUCAAACAAGGCAAG AAGUCAGGCCGGAUUAAGCCAUAGCACGGUAAGAGCUAUGCUGC CUGUGAGCCCCGUCCAAGGACGUAAAAUGAAGUCAGGCCGAAAG CCACGGUUCGAGCAAGCCGUGCUGCCUGUAGCUCCAUCGUGGGG AUGUAAAAACCCGGGAGGCUGCAAACCAUGGAAGCUGUACGCAU GGGGUAGCAGACUAGUGGUUAGAGGAGACCCCUCCCAAGACACA ACGCAGCAGCGGGGCCCAACACCAGGGGAAGCUGUACCCUGGUG GUAAGGACUAGAGGUUAGAGGAGACCCCCCGCACAACAACAAAC AGCAUAUUGACGCUGGGAGAGACCAGAGAUCCUGCUGUCUCUAC AGCAUCAUUCCAGGCACAGAACGCCAAAAAAUGGAAUGGUGCUG UUGAAUCAACAGGUUCU DEN-2 AGUUGUUAGUCUACGUGGACCGACAAAGACAGAUUCUUUGAGG 241 (NC_001474.2) GAGCUAAGCUCAACGUAGUUCUAACAGUUUUUUAAUUAGAGAG CAGAUCUCUGAUGAAUAACCAACGGAAAAAGGCGAAAAACACGC CUUUCAAUAUGCUGAAACGCGAGAGAAACCGCGUGUCGACUGUG CAACAGCUGACAAAGAGAUUCUCACUUGGAAUGCUGCAGGGACG AGGACCAUUAAAACUGUUCAUGGCCCUGGUGGCGUUCCUUCGUU UCCUAACAAUCCCACCAACAGCAGGGAUAUUGAAGAGAUGGGGA ACAAUUAAAAAAUCAAAAGCUAUUAAUGUUUUGAGAGGGUUCA GGAAAGAGAUUGGAAGGAUGCUGAACAUCUUGAAUAGGAGACG CAGAUCUGCAGGCAUGAUCAUUAUGCUGAUUCCAACAGUGAUGG CGUUCCAUUUAACCACACGUAACGGAGAACCACACAUGAUCGUC AGCAGACAAGAGAAAGGGAAAAGUCUUCUGUUUAAAACAGAGG AUGGCGUGAACAUGUGUACCCUCAUGGCCAUGGACCUUGGUGAA UUGUGUGAAGACACAAUCACGUACAAGUGUCCCCUUCUCAGGCA GAAUGAGCCAGAAGACAUAGACUGUUGGUGCAACUCUACGUCCA CGUGGGUAACUUAUGGGACGUGUACCACCAUGGGAGAACAUAG AAGAGAAAAAAGAUCAGUGGCACUCGUUCCACAUGUGGGAAUG GGACUGGAGACACGAACUGAAACAUGGAUGUCAUCAGAAGGGG CCUGGAAACAUGUCCAGAGAAUUGAAACUUGGAUCUUGAGACA UCCAGGCUUCACCAUGAUGGCAGCAAUCCUGGCAUACACCAUAG GAACGACACAUUUCCAAAGAGCCCUGAUUUUCAUCUUACUGACA GCUGUCACUCCUUCAAUGACAAUGCGUUGCAUAGGAAUGUCAAA UAGAGACUUUGUGGAAGGGGUUUCAGGAGGAAGCUGGGUUGAC AUAGUCUUAGAACAUGGAAGCUGUGUGACGACGAUGGCAAAAA ACAAACCAACAUUGGAUUUUGAACUGAUAAAAACAGAAGCCAA ACAGCCUGCCACCCUAAGGAAGUACUGUAUAGAGGCAAAGCUAA CCAACACAACAACAGAAUCUCGCUGCCCAACACAAGGGGAACCC AGCCUAAAUGAAGAGCAGGACAAAAGGUUCGUCUGCAAACACUC CAUGGUAGACAGAGGAUGGGGAAAUGGAUGUGGACUAUUUGGA AAGGGAGGCAUUGUGACCUGUGCUAUGUUCAGAUGCAAAAAGA ACAUGGAAGGAAAAGUUGUGCAACCAGAAAACUUGGAAUACAC CAUUGUGAUAACACCUCACUCAGGGGAAGAGCAUGCAGUCGGAA AUGACACAGGAAAACAUGGCAAGGAAAUCAAAAUAACACCACAG AGUUCCAUCACAGAAGCAGAAUUGACAGGUUAUGGCACUGUCAC AAUGGAGUGCUCUCCAAGAACGGGCCUCGACUUCAAUGAGAUGG UGUUGCUGCAGAUGGAAAAUAAAGCUUGGCUGGUGCACAGGCA AUGGUUCCUAGACCUGCCGUUACCAUGGUUGCCCGGAGCGGACA CACAAGGGUCAAAUUGGAUACAGAAAGAGACAUUGGUCACUUU CAAAAAUCCCCAUGCGAAGAAACAGGAUGUUGUUGUUUUAGGA UCCCAAGAAGGGGCCAUGCACACAGCACUUACAGGGGCCACAGA AAUCCAAAUGUCAUCAGGAAACUUACUCUUCACAGGACAUCUCA AGUGCAGGCUGAGAAUGGACAAGCUACAGCUCAAAGGAAUGUC AUACUCUAUGUGCACAGGAAAGUUUAAAGUUGUGAAGGAAAUA GCAGAAACACAACAUGGAACAAUAGUUAUCAGAGUGCAAUAUG AAGGGGACGGCUCUCCAUGCAAGAUCCCUUUUGAGAUAAUGGAU UUGGAAAAAAGACAUGUCUUAGGUCGCCUGAUUACAGUCAACCC AAUUGUGACAGAAAAAGAUAGCCCAGUCAACAUAGAAGCAGAA CCUCCAUUCGGAGACAGCUACAUCAUCAUAGGAGUAGAGCCGGG ACAACUGAAGCUCAACUGGUUUAAGAAAGGAAGUUCUAUCGGCC AAAUGUUUGAGACAACAAUGAGGGGGGCGAAGAGAAUGGCCAU UUUAGGUGACACAGCCUGGGAUUUUGGAUCCUUGGGAGGAGUG UUUACAUCUAUAGGAAAGGCUCUCCACCAAGUCUUUGGAGCAAU CUAUGGAGCUGCCUUCAGUGGGGUUUCAUGGACUAUGAAAAUCC UCAUAGGAGUCAUUAUCACAUGGAUAGGAAUGAAUUCACGCAG CACCUCACUGUCUGUGACACUAGUAUUGGUGGGAAUUGUGACAC UGUAUUUGGGAGUCAUGGUGCAGGCCGAUAGUGGUUGCGUUGU GAGCUGGAAAAACAAAGAACUGAAAUGUGGCAGUGGGAUUUUC AUCACAGACAACGUGCACACAUGGACAGAACAAUACAAGUUCCA ACCAGAAUCCCCUUCAAAACUAGCUUCAGCUAUCCAGAAAGCCC AUGAAGAGGGCAUUUGUGGAAUCCGCUCAGUAACAAGACUGGA GAAUCUGAUGUGGAAACAAAUAACACCAGAAUUGAAUCACAUU CUAUCAGAAAAUGAGGUGAAGUUAACUAUUAUGACAGGAGACA UCAAAGGAAUCAUGCAGGCAGGAAAACGAUCUCUGCGGCCUCAG CCCACUGAGCUGAAGUAUUCAUGGAAAACAUGGGGCAAAGCAAA AAUGCUCUCUACAGAGUCUCAUAACCAGACCUUUCUCAUUGAUG GCCCCGAAACAGCAGAAUGCCCCAACACAAAUAGAGCUUGGAAU UCGUUGGAAGUUGAAGACUAUGGCUUUGGAGUAUUCACCACCA AUAUAUGGCUAAAAUUGAAAGAAAAACAGGAUGUAUUCUGCGA CUCAAAACUCAUGUCAGCGGCCAUAAAAGACAACAGAGCCGUCC AUGCCGAUAUGGGUUAUUGGAUAGAAAGUGCACUCAAUGACAC AUGGAAGAUAGAGAAAGCCUCUUUCAUUGAAGUUAAAAACUGC CACUGGCCAAAAUCACACACCCUCUGGAGCAAUGGAGUGCUAGA AAGUGAGAUGAUAAUUCCAAAGAAUCUCGCUGGACCAGUGUCUC AACACAACUAUAGACCAGGCUACCAUACACAAAUAACAGGACCA UGGCAUCUAGGUAAGCUUGAGAUGGACUUUGAUUUCUGUGAUG GAACAACAGUGGUAGUGACUGAGGACUGCGGAAAUAGAGGACC CUCUUUGAGAACAACCACUGCCUCUGGAAAACUCAUAACAGAAU GGUGCUGCCGAUCUUGCACAUUACCACCGCUAAGAUACAGAGGU GAGGAUGGGUGCUGGUACGGGAUGGAAAUCAGACCAUUGAAGG AGAAAGAAGAGAAUUUGGUCAACUCCUUGGUCACAGCUGGACA UGGGCAGGUCGACAACUUUUCACUAGGAGUCUUGGGAAUGGCA UUGUUCCUGGAGGAAAUGCUUAGGACCCGAGUAGGAACGAAAC AUGCAAUACUACUAGUUGCAGUUUCUUUUGUGACAUUGAUCAC AGGGAACAUGUCCUUUAGAGACCUGGGAAGAGUGAUGGUUAUG GUAGGCGCCACUAUGACGGAUGACAUAGGUAUGGGCGUGACUU AUCUUGCCCUACUAGCAGCCUUCAAAGUCAGACCAACUUUUGCA GCUGGACUACUCUUGAGAAAGCUGACCUCCAAGGAAUUGAUGAU GACUACUAUAGGAAUUGUACUCCUCUCCCAGAGCACCAUACCAG AGACCAUUCUUGAGUUGACUGAUGCGUUAGCCUUAGGCAUGAU GGUCCUCAAAAUGGUGAGAAAUAUGGAAAAGUAUCAAUUGGCA GUGACUAUCAUGGCUAUCUUGUGCGUCCCAAACGCAGUGAUAUU ACAAAACGCAUGGAAAGUGAGUUGCACAAUAUUGGCAGUGGUG UCCGUUUCCCCACUGCUCUUAACAUCCUCACAGCAAAAAACAGA UUGGAUACCAUUAGCAUUGACGAUCAAAGGUCUCAAUCCAACAG CUAUUUUUCUAACAACCCUCUCAAGAACCAGCAAGAAAAGGAGC UGGCCAUUAAAUGAGGCUAUCAUGGCAGUCGGGAUGGUGAGCA UUUUAGCCAGUUCUCUCCUAAAAAAUGAUAUUCCCAUGACAGGA CCAUUAGUGGCUGGAGGGCUCCUCACUGUGUGCUACGUGCUCAC UGGACGAUCGGCCGAUUUGGAACUGGAGAGAGCAGCCGAUGUCA AAUGGGAAGACCAGGCAGAGAUAUCAGGAAGCAGUCCAAUCCUG UCAAUAACAAUAUCAGAAGAUGGUAGCAUGUCGAUAAAAAAUG AAGAGGAAGAACAAACACUGACCAUACUCAUUAGAACAGGAUU GCUGGUGAUCUCAGGACUUUUUCCUGUAUCAAUACCAAUCACGG CAGCAGCAUGGUACCUGUGGGAAGUGAAGAAACAACGGGCCGGA GUAUUGUGGGAUGUUCCUUCACCCCCACCCAUGGGAAAGGCUGA ACUGGAAGAUGGAGCCUAUAGAAUUAAGCAAAAAGGGAUUCUU GGAUAUUCCCAGAUCGGAGCCGGAGUUUACAAAGAAGGAACAU UCCAUACAAUGUGGCAUGUCACACGUGGCGCUGUUCUAAUGCAU AAAGGAAAGAGGAUUGAACCAUCAUGGGCGGACGUCAAGAAAG ACCUAAUAUCAUAUGGAGGAGGCUGGAAGUUAGAAGGAGAAUG GAAGGAAGGAGAAGAAGUCCAGGUAUUGGCACUGGAGCCUGGA AAAAAUCCAAGAGCCGUCCAAACGAAACCUGGUCUUUUCAAAAC CAACGCCGGAACAAUAGGUGCUGUAUCUCUGGACUUUUCUCCUG GAACGUCAGGAUCUCCAAUUAUCGACAAAAAAGGAAAAGUUGU GGGUCUUUAUGGUAAUGGUGUUGUUACAAGGAGUGGAGCAUAU GUGAGUGCUAUAGCCCAGACUGAAAAAAGCAUUGAAGACAACCC AGAGAUCGAAGAUGACAUUUUCCGAAAGAGAAGACUGACCAUC AUGGACCUCCACCCAGGAGCGGGAAAGACGAAGAGAUACCUUCC GGCCAUAGUCAGAGAAGCUAUAAAACGGGGUUUGAGAACAUUA AUCUUGGCCCCCACUAGAGUUGUGGCAGCUGAAAUGGAGGAAGC CCUUAGAGGACUUCCAAUAAGAUACCAGACCCCAGCCAUCAGAG CUGAGCACACCGGGCGGGAGAUUGUGGACCUAAUGUGUCAUGCC ACAUUUACCAUGAGGCUGCUAUCACCAGUUAGAGUGCCAAACUA CAACCUGAUUAUCAUGGACGAAGCCCAUUUCACAGACCCAGCAA GUAUAGCAGCUAGAGGAUACAUCUCAACUCGAGUGGAGAUGGG UGAGGCAGCUGGGAUUUUUAUGACAGCCACUCCCCCGGGAAGCA GAGACCCAUUUCCUCAGAGCAAUGCACCAAUCAUAGAUGAAGAA AGAGAAAUCCCUGAACGUUCGUGGAAUUCCGGACAUGAAUGGG UCACGGAUUUUAAAGGGAAGACUGUUUGGUUCGUUCCAAGUAU AAAAGCAGGAAAUGAUAUAGCAGCUUGCCUGAGGAAAAAUGGA AAGAAAGUGAUACAACUCAGUAGGAAGACCUUUGAUUCUGAGU AUGUCAAGACUAGAACCAAUGAUUGGGACUUCGUGGUUACAAC UGACAUUUCAGAAAUGGGUGCCAAUUUCAAGGCUGAGAGGGUU AUAGACCCCAGACGCUGCAUGAAACCAGUCAUACUAACAGAUGG UGAAGAGCGGGUGAUUCUGGCAGGACCUAUGCCAGUGACCCACU CUAGUGCAGCACAAAGAAGAGGGAGAAUAGGAAGAAAUCCAAA AAAUGAGAAUGACCAGUACAUAUACAUGGGGGAACCUCUGGAA AAUGAUGAAGACUGUGCACACUGGAAAGAAGCUAAAAUGCUCC UAGAUAACAUCAACACGCCAGAAGGAAUCAUUCCUAGCAUGUUC GAACCAGAGCGUGAAAAGGUGGAUGCCAUUGAUGGCGAAUACC GCUUGAGAGGAGAAGCAAGGAAAACCUUUGUAGACUUAAUGAG AAGAGGAGACCUACCAGUCUGGUUGGCCUACAGAGUGGCAGCUG AAGGCAUCAACUACGCAGACAGAAGGUGGUGUUUUGAUGGAGU CAAGAACAACCAAAUCCUAGAAGAAAACGUGGAAGUUGAAAUC UGGACAAAAGAAGGGGAAAGGAAGAAAUUGAAACCCAGAUGGU UGGAUGCUAGGAUCUAUUCUGACCCACUGGCGCUAAAAGAAUUU AAGGAAUUUGCAGCCGGAAGAAAGUCUCUGACCCUGAACCUAAU CACAGAAAUGGGUAGGCUCCCAACCUUCAUGACUCAGAAGGCAA GAGACGCACUGGACAACUUAGCAGUGCUGCACACGGCUGAGGCA GGUGGAAGGGCGUACAACCAUGCUCUCAGUGAACUGCCGGAGAC CCUGGAGACAUUGCUUUUACUGACACUUCUGGCUACAGUCACGG GAGGGAUCUUUUUAUUCUUGAUGAGCGGAAGGGGCAUAGGGAA GAUGACCCUGGGAAUGUGCUGCAUAAUCACGGCUAGCAUCCUCC UAUGGUACGCACAAAUACAGCCACACUGGAUAGCAGCUUCAAUA AUACUGGAGUUUUUUCUCAUAGUUUUGCUUAUUCCAGAACCUG AAAAACAGAGAACACCCCAAGACAACCAACUGACCUACGUUGUC AUAGCCAUCCUCACAGUGGUGGCCGCAACCAUGGCAAACGAGAU GGGUUUCCUAGAAAAAACGAAGAAAGAUCUCGGAUUGGGAAGC AUUGCAACCCAGCAACCCGAGAGCAACAUCCUGGACAUAGAUCU ACGUCCUGCAUCAGCAUGGACGCUGUAUGCCGUGGCCACAACAU UUGUUACACCAAUGUUGAGACAUAGCAUUGAAAAUUCCUCAGU GAAUGUGUCCCUAACAGCUAUAGCCAACCAAGCCACAGUGUUAA UGGGUCUCGGGAAAGGAUGGCCAUUGUCAAAGAUGGACAUCGG AGUUCCCCUUCUCGCCAUUGGAUGCUACUCACAAGUCAACCCCA UAACUCUCACAGCAGCUCUUUUCUUAUUGGUAGCACAUUAUGCC AUCAUAGGGCCAGGACUCCAAGCAAAAGCAACCAGAGAAGCUCA GAAAAGAGCAGCGGCGGGCAUCAUGAAAAACCCAACUGUCGAUG GAAUAACAGUGAUUGACCUAGAUCCAAUACCUUAUGAUCCAAAG UUUGAAAAGCAGUUGGGACAAGUAAUGCUCCUAGUCCUCUGCGU GACUCAAGUAUUGAUGAUGAGGACUACAUGGGCUCUGUGUGAG GCUUUAACCUUAGCUACCGGGCCCAUCUCCACAUUGUGGGAAGG AAAUCCAGGGAGGUUUUGGAACACUACCAUUGCGGUGUCAAUG GCUAACAUUUUUAGAGGGAGUUACUUGGCCGGAGCUGGACUUC UCUUUUCUAUUAUGAAGAACACAACCAACACAAGAAGGGGAACU GGCAACAUAGGAGAGACGCUUGGAGAGAAAUGGAAAAGCCGAU UGAACGCAUUGGGAAAAAGUGAAUUCCAGAUCUACAAGAAAAG UGGAAUCCAGGAAGUGGAUAGAACCUUAGCAAAAGAAGGCAUU AAAAGAGGAGAAACGGACCAUCACGCUGUGUCGCGAGGCUCAGC AAAACUGAGAUGGUUCGUUGAGAGAAACAUGGUCACACCAGAA GGGAAAGUAGUGGACCUCGGUUGUGGCAGAGGAGGCUGGUCAU ACUAUUGUGGAGGACUAAAGAAUGUAAGAGAAGUCAAAGGCCU AACAAAAGGAGGACCAGGACACGAAGAACCCAUCCCCAUGUCAA CAUAUGGGUGGAAUCUAGUGCGUCUUCAAAGUGGAGUUGACGU UUUCUUCAUCCCGCCAGAAAAGUGUGACACAUUAUUGUGUGACA UAGGGGAGUCAUCACCAAAUCCCACAGUGGAAGCAGGACGAACA CUCAGAGUCCUUAACUUAGUAGAAAAUUGGUUGAACAACAACAC UCAAUUUUGCAUAAAGGUUCUCAACCCAUAUAUGCCCUCAGUCA UAGAAAAAAUGGAAGCACUACAAAGGAAAUAUGGAGGAGCCUU AGUGAGGAAUCCACUCUCACGAAACUCCACACAUGAGAUGUACU GGGUAUCCAAUGCUUCCGGGAACAUAGUGUCAUCAGUGAACAUG AUUUCAAGGAUGUUGAUCAACAGAUUUACAAUGAGAUACAAGA AAGCCACUUACGAGCCGGAUGUUGACCUCGGAAGCGGAACCCGU AACAUCGGGAUUGAAAGUGAGAUACCAAACCUAGAUAUAAUUG GGAAAAGAAUAGAAAAAAUAAAGCAAGAGCAUGAAACAUCAUG GCACUAUGACCAAGACCACCCAUACAAAACGUGGGCAUACCAUG GUAGCUAUGAAACAAAACAGACUGGAUCAGCAUCAUCCAUGGUC AACGGAGUGGUCAGGCUGCUGACAAAACCUUGGGACGUCGUCCC CAUGGUGACACAGAUGGCAAUGACAGACACGACUCCAUUUGGAC AACAGCGCGUUUUUAAAGAGAAAGUGGACACGAGAACCCAAGA ACCGAAAGAAGGCACGAAGAAACUAAUGAAAAUAACAGCAGAG UGGCUUUGGAAAGAAUUAGGGAAGAAAAAGACACCCAGGAUGU GCACCAGAGAAGAAUUCACAAGAAAGGUGAGAAGCAAUGCAGCC UUGGGGGCCAUAUUCACUGAUGAGAACAAGUGGAAGUCGGCAC GUGAGGCUGUUGAAGAUAGUAGGUUUUGGGAGCUGGUUGACAA GGAAAGGAAUCUCCAUCUUGAAGGAAAGUGUGAAACAUGUGUG UACAACAUGAUGGGAAAAAGAGAGAAGAAGCUAGGGGAAUUCG GCAAGGCAAAAGGCAGCAGAGCCAUAUGGUACAUGUGGCUUGG AGCACGCUUCUUAGAGUUUGAAGCCCUAGGAUUCUUAAAUGAA GAUCACUGGUUCUCCAGAGAGAACUCCCUGAGUGGAGUGGAAGG AGAAGGGCUGCACAAGCUAGGUUACAUUCUAAGAGACGUGAGC AAGAAAGAGGGAGGAGCAAUGUAUGCCGAUGACACCGCAGGAU GGGAUACAAGAAUCACACUAGAAGACCUAAAAAAUGAAGAAAU GGUAACAAACCACAUGGAAGGAGAACACAAGAAACUAGCCGAGG CCAUUUUCAAACUAACGUACCAAAACAAGGUGGUGCGUGUGCAA AGACCAACACCAAGAGGCACAGUAAUGGACAUCAUAUCGAGAAG AGACCAAAGAGGUAGUGGACAAGUUGGCACCUAUGGACUCAAU ACUUUCACCAAUAUGGAAGCCCAACUAAUCAGACAGAUGGAGGG AGAAGGAGUCUUUAAAAGCAUUCAGCACCUAACAAUCACAGAAG AAAUCGCUGUGCAAAACUGGUUAGCAAGAGUGGGGCGCGAAAG GUUAUCAAGAAUGGCCAUCAGUGGAGAUGAUUGUGUUGUGAAA CCUUUAGAUGACAGGUUCGCAAGCGCUUUAACAGCUCUAAAUGA CAUGGGAAAGAUUAGGAAAGACAUACAACAAUGGGAACCUUCA AGAGGAUGGAAUGAUUGGACACAAGUGCCCUUCUGUUCACACCA UUUCCAUGAGUUAAUCAUGAAAGACGGUCGCGUACUCGUUGUUC CAUGUAGAAACCAAGAUGAACUGAUUGGCAGAGCCCGAAUCUCC CAAGGAGCAGGGUGGUCUUUGCGGGAGACGGCCUGUUUGGGGA AGUCUUACGCCCAAAUGUGGAGCUUGAUGUACUUCCACAGACGC GACCUCAGGCUGGCGGCAAAUGCUAUUUGCUCGGCAGUACCAUC ACAUUGGGUUCCAACAAGUCGAACAACCUGGUCCAUACAUGCUA AACAUGAAUGGAUGACAACGGAAGACAUGCUGACAGUCUGGAA CAGGGUGUGGAUUCAAGAAAACCCAUGGAUGGAAGACAAAACU CCAGUGGAAUCAUGGGAGGAAAUCCCAUACUUGGGGAAAAGAG AAGACCAAUGGUGCGGCUCAUUGAUUGGGUUAACAAGCAGGGCC ACCUGGGCAAAGAACAUCCAAGCAGCAAUAAAUCAAGUUAGAUC CCUUAUAGGCAAUGAAGAAUACACAGAUUACAUGCCAUCCAUGA AAAGAUUCAGAAGAGAAGAGGAAGAAGCAGGAGUUCUGUGGUA GAAAGCAAAACUAACAUGAAACAAGGCUAGAAGUCAGGUCGGA UUAAGCCAUAGUACGGAAAAAACUAUGCUACCUGUGAGCCCCGU CCAAGGACGUUAAAAGAAGUCAGGCCAUCAUAAAUGCCAUAGCU UGAGUAAACUAUGCAGCCUGUAGCUCCACCUGAGAAGGUGUAAA AAAUCCGGGAGGCCACAAACCAUGGAAGCUGUACGCAUGGCGUA GUGGACUAGCGGUUAGAGGAGACCCCUCCCUUACAAAUCGCAGC AACAAUGGGGGCCCAAGGCGAGAUGAAGCUGUAGUCUCGCUGGA AGGACUAGAGGUUAGAGGAGACCCCCCCGAAACAAAAAACAGCA UAUUGACGCUGGGAAAGACCAGAGAUCCUGCUGUCUCCUCAGCA UCAUUCCAGGCACAGAACGCCAGAAAAUGGAAUGGUGCUGUUGA AUCAACAGGUUCU DEN-3 AGUUGUUAGUCUACGUGGACCGACAAGAACAGUUUCGACUCGGA 242 (NC_001475.2) AGCUUGCUUAACGUAGUGCUGACAGUUUUUUAUUAGAGAGCAG AUCUCUGAUGAACAACCAACGGAAGAAGACGGGAAAACCGUCUA UCAAUAUGCUGAAACGCGUGAGAAACCGUGUGUCAACUGGAUCA CAGUUGGCGAAGAGAUUCUCAAAAGGACUGCUGAACGGCCAGGG ACCAAUGAAAUUGGUUAUGGCGUUCAUAGCUUUCCUCAGAUUUC UAGCCAUUCCACCAACAGCAGGAGUCUUGGCUAGAUGGGGAACC UUCAAGAAGUCGGGGGCCAUUAAGGUCCUGAAAGGCUUCAAGA AGGAGAUCUCAAACAUGCUGAGCAUAAUCAACCAACGGAAAAAG ACAUCGCUCUGUCUCAUGAUGAUAUUGCCAGCAGCACUUGCUUU CCACUUGACUUCACGAGAUGGAGAGCCGCGCAUGAUUGUGGGGA AGAAUGAAAGAGGUAAAUCCCUACUUUUUAAGACAGCCUCUGG AAUCAACAUGUGCACACUCAUAGCCAUGGAUUUGGGAGAGAUG UGUGAUGACACGGUCACUUACAAAUGCCCCCACAUUACCGAAGU GGAACCUGAAGACAUUGACUGCUGGUGCAACCUUACAUCAACAU GGGUGACUUAUGGAACGUGCAAUCAAGCUGGAGAGCAUAGACG CGACAAGAGAUCAGUGGCGUUAGCUCCCCAUGUCGGCAUGGGAC UGGACACACGCACCCAAACCUGGAUGUCGGCUGAAGGAGCUUGG AGACAAGUCGAGAAGGUAGAGACAUGGGCCCUUAGGCACCCAGG GUUCACCAUACUAGCCCUAUUUCUCGCCCAUUACAUAGGCACUU CCCUGACCCAGAAGGUGGUUAUUUUCAUAUUAUUAAUGCUGGUC ACCCCAUCCAUGACAAUGAGAUGUGUGGGAGUAGGAAACAGAG AUUUUGUGGAAGGGCUAUCAGGAGCUACGUGGGUUGACGUGGU GCUCGAGCACGGGGGGUGUGUGACUACCAUGGCUAAGAACAAGC CCACGCUGGAUAUAGAGCUUCAGAAGACCGAGGCCACCCAACUG GCGACCCUAAGGAAGCUAUGCAUUGAGGGGAAAAUUACCAACAU AACAACUGACUCAAGAUGUCCUACCCAAGGGGAAGCGGUUUUGC CUGAGGAGCAGGACCAGAACUACGUGUGUAAGCAUACAUACGUA GACAGAGGUUGGGGGAACGGUUGUGGUUUGUUUGGCAAAGGAA GCUUGGUAACAUGUGCGAAAUUUCAAUGCCUGGAACCAAUAGA GGGAAAAGUGGUGCAAUAUGAGAACCUCAAAUACACCGUCAUCA UUACAGUGCACACAGGAGACCAACACCAGGUGGGAAAUGAAACG CAAGGAGUCACGGCUGAGAUAACACCUCAGGCAUCAACCACUGA AGCCAUCUUGCCUGAAUAUGGAACCCUUGGGCUAGAAUGCUCAC CACGGACAGGUUUGGAUUUCAAUGAAAUGAUCUUACUAACAAU GAAGAACAAAGCAUGGAUGGUACAUAGACAAUGGUUCUUUGAC CUACCUCUACCAUGGGCAUCAGGAGCUACAACAGAAACACCAAC CUGGAACAGGAAGGAGCUUCUUGUGACAUUCAAAAACGCACAUG CGAAAAAACAAGAAGUAGUUGUCCUUGGAUCGCAAGAGGGAGC AAUGCAUACCGCACUGACAGGAGCUACAGAAAUCCAAAACUCAG GAGGCACAAGCAUUUUCGCGGGGCACUUAAAAUGUAGACUUAA GAUGGACAAAUUGGAACUCAAGGGGAUGAGCUAUGCAAUGUGC ACGAAUACCUUUGUGUUGAAGAAAGAAGUCUCAGAAACGCAGC ACGGGACAAUACUCAUUAAGGUUGAGUACAAAGGGGAAGAUGC ACCUUGCAAGAUUCCCUUUUCCACAGAGGAUGGACAAGGGAAAG CUCAUAAUGGCAGACUGAUCACAGCCAACCCUGUGGUGACUAAG AAGGAGGAGCCUGUCAAUAUUGAGGCUGAACCUCCUUUUGGGG AAAGCAAUAUAGUAAUUGGAAUUGGAGACAACGCCUUGAAAAU CAACUGGUACAAGAAGGGGAGCUCGAUUGGGAAGAUGUUCGAG GCCACUGAAAGGGGUGCAAGGCGCAUGGCCAUCUUGGGAGACAC AGCUUGGGACUUUGGAUCAGUGGGUGGUGUUCUGAACUCAUUA GGCAAAAUGGUGCACCAAAUAUUUGGAAGUGCUUAUACAGCCCU GUUCAGUGGAGUCUCUUGGGUGAUGAAAAUUGGAAUAGGUGUC CUCUUGACUUGGAUAGGGUUGAAUUCAAAAAACACAUCCAUGUC AUUUUCAUGCAUUGCGAUAGGAAUCAUUACACUCUAUCUGGGA GCUGUGGUACAAGCUGACAUGGGGUGUGUCAUAAACUGGAAGG GCAAAGAACUCAAAUGUGGAAGCGGAAUUUUCGUCACCAAUGA GGUCCAUACCUGGACAGAGCAAUACAAAUUCCAAGCAGACUCCC CAAAAAGAUUGGCAACAGCCAUUGCAGGCGCCUGGGAGAAUGGA GUGUGUGGAAUUAGGUCAACAACCAGAAUGGAGAAUCUCUUGU GGAAGCAAAUAGCCAAUGAACUGAACUACAUAUUAUGGGAAAA CAAUAUCAAAUUAACGGUAGUUGUGGGCGAUACACUUGGGGUC UUAGAGCAAGGGAAAAGAACACUAACACCACAACCCAUGGAGCU AAAAUACUCAUGGAAAACGUGGGGAAAGGCAAAAAUAGUGACA GCUGAAACACAAAAUUCCUCUUUCAUAAUAGACGGGCCAAACAC ACCGGAGUGUCCAAGUGCCUCAAGAGCAUGGAAUGUGUGGGAG GUGGAAGAUUACGGGUUCGGAGUCUUCACAACCAACAUAUGGCU GAAACUCCGAGAGGUCUACACCCAACUAUGUGACCAUAGGCUAA UGUCGGCAGCUGUCAAGGAUGAGAGGGCCGUGCAUGCCGACAUG GGCUACUGGAUAGAAAGCCAAAAGAAUGGAAGUUGGAAGCUAG AAAAAGCAUCCCUCAUAGAGGUAAAAACCUGCACAUGGCCAAAA UCACACACUCUCUGGACUAAUGGUGUGCUAGAGAGUGACAUGAU CAUCCCAAAGAGUCUAGCUGGUCCUAUCUCACAACACAACUACA GGCCCGGGUACCACACCCAAACGGCAGGACCCUGGCACUUAGGA AAAUUGGAGCUGGACUUCAACUACUGUGAAGGAACAACAGUUG UCAUCACAGAAAGCUGUGGGACAAGAGGCCCAUCAUUGAGAACA ACAACAGUGUCAGGGAAGUUGAUACACGAAUGGUGUUGCCGCUC GUGCACACUUCCCCCCCUGCGAUACAUGGGAGAAGACGGCUGCU GGUAUGGCAUGGAAAUCAGACCCAUCAGUGAGAAAGAAGAGAA CAUGGUAAAGUCUUUAGUCUCAGCGGGAAGUGGAAAGGUGGAC AACUUCACAAUGGGUGUCUUGUGUUUGGCAAUCCUCUUUGAAG AGGUGUUGAGAGGAAAAUUUGGGAAGAAACACAUGAUUGCAGG GGUUUUCUUUACGUUUGUGCUCCUUCUCUCAGGGCAAAUAACAU GGAGAGACAUGGCGCACACACUAAUAAUGAUCGGGUCCAACGCC UCUGACAGGAUGGGAAUGGGCGUCACCUACCUAGCUCUAAUUGC AACAUUUAAAAUCCAGCCAUUCUUGGCUUUGGGAUUUUUCCUAA GAAAGCUGACAUCUAGAGAAAAUUUAUUGUUAGGAGUUGGGUU GGCCAUGGCAACAACGUUACAACUGCCAGAGGACAUUGAACAAA UGGCAAAUGGAGUCGCUCUGGGGCUCAUGGCUCUUAAACUGAUA ACACAAUUUGAAACAUACCAAUUGUGGACGGCAUUAGUCUCCUU AACGUGUUCAAACACAAUUUUUACGUUGACUGUUGCCUGGAGA ACAGCCACUCUGAUUUUGGCCGGAGUUUCGCUUUUACCAGUGUG CCAGUCUUCAAGCAUGAGGAAAACAGAUUGGCUCCCAAUGACAG UGGCAGCUAUGGGAGUUCCACCCCUUCCACUUUUUAUUUUUAGC UUGAAAGACACACUCAAAAGGAGAAGCUGGCCACUGAAUGAAG GGGUGAUGGCUGUUGGGCUUGUGAGCAUUCUGGCCAGUUCUCUC CUUAGAAAUGAUGUGCCCAUGGCUGGACCAUUAGUGGCCGGGGG CUUGCUGAUAGCGUGCUACGUCAUAACUGGCACGUCAGCGGACC UCACUGUAGAAAAAGCCCCAGAUGUAACAUGGGAGGAAGAGGC UGAGCAGACAGGAGUGUCCCACAACUUAAUGAUCACAGUUGAUG AUGAUGGAACAAUGAGAAUAAAAGAUGAUGAGACUGAGAACAU CCUAACAGUGCUUUUAAAAACAGCAUUACUAAUAGUAUCAGGCA UUUUUCCAUACUCCAUACCCGCAACAUUGUUGGUCUGGCACACU UGGCAAAAACAAACCCAAAGAUCCGGCGUUUUAUGGGACGUACC CAGCCCCCCAGAGACACAGAAAGCAGAACUGGAAGAAGGGGUUU AUAGGAUCAAACAGCAAGGAAUUUUUGGGAAAACCCAAGUAGG GGUUGGAGUACAGAAAGAAGGAGUCUUCCACACCAUGUGGCACG UCACAAGAGGGGCAGUGUUGACACAUAAUGGGAAAAGACUGGA ACCAAACUGGGCUAGUGUGAAAAAAGAUCUGAUUUCAUAUGGA GGAGGAUGGAGACUGAGCGCACAAUGGCAAAAGGGGGAGGAGG UGCAGGUUAUUGCCGUAGAGCCAGGGAAGAACCCAAAGAACUUU CAAACCACGCCAGGCACUUUCCAGACUACUACAGGGGAAAUAGG AGCAAUUGCACUGGAUUUCAAGCCUGGAACUUCAGGAUCUCCUA UCAUAAAUAGAGAGGGAAAGGUAGUGGGACUGUAUGGCAAUGG AGUGGUUACAAAGAAUGGUGGCUAUGUCAGCGGAAUAGCGCAA ACAAAUGCAGAACCAGAUGGACCGACACCAGAGUUGGAAGAAGA GAUGUUCAAAAAGCGAAACCUGACCAUAAUGGAUCUUCAUCCUG GGUCAGGAAAGACACGGAAAUACCUUCCAGCUAUUGUCAGAGAG GCAAUCAAGAGACGUUUAAGAACCUUAAUUUUGGCACCGACAAG GGUGGUUGCAGCUGAGAUGGAAGAAGCAUUGAAAGGGCUCCCA AUAAGGUACCAAACAACAGCAACAAAAUCUGAACACACAGGAAG AGAGAUUGUUGAUCUAAUGUGCCACGCAACGUUCACAAUGCGUU UGCUGUCACCAGUUAGGGUUCCAAAUUACAACUUGAUAAUAAU GGAUGAGGCCCAUUUCACAGACCCAGCCAGUAUAGCGGCUAGAG GGUACAUAUCAACUCGUGUUGGAAUGGGAGAGGCAGCCGCAAUC UUCAUGACAGCAACACCCCCUGGAACAGCUGAUGCCUUUCCUCA GAGCAACGCUCCAAUUCAAGAUGAAGAAAGGGACAUACCAGAAC GCUCAUGGAAUUCAGGCAAUGAAUGGAUUACCGACUUCGCUGGG AAAACGGUGUGGUUUGUCCCUAGCAUUAAAGCCGGAAAUGACA UAGCAAACUGCUUGCGAAAAAACGGGAAAAAAGUCAUUCAACU UAGUAGGAAGACUUUUGACACAGAAUAUCAGAAGACUAAACUG AAUGAUUGGGACUUUGUGGUGACAACUGACAUUUCAGAAAUGG GGGCCAAUUUCAAAGCAGAUAGAGUGAUCGACCCAAGAAGAUG UCUCAAACCAGUGAUCUUGACAGAUGGACCAGAGCGGGUGAUCC UGGCCGGACCAAUGCCAGUCACCGCGGCGAGUGCUGCGCAAAGG AGAGGGAGAGUUGGCAGGAACCCACAAAAAGAGAAUGACCAGU ACAUAUUCACGGGCCAGCCUCUCAACAAUGAUGAAGACCAUGCU CACUGGACAGAAGCAAAAAUGCUGCUGGACAACAUCAACACACC AGAAGGGAUUAUACCAGCUCUCUUUGAACCAGAAAGGGAGAAG UCAGCCGCCAUAGACGGUGAGUAUCGCCUGAAGGGUGAGUCCAG GAAGACUUUCGUGGAACUCAUGAGGAGGGGUGACCUUCCAGUU UGGUUAGCCCAUAAAGUAGCAUCAGAAGGAAUCAAAUACACAG AUAGAAAAUGGUGCUUUGAUGGGCAACGCAAUAAUCAAAUUUU AGAGGAGAACAUGGAUGUGGAAAUUUGGACAAAGGAAGGAGAA AAGAAAAAAUUGAGACCUAGGUGGCUUGAUGCCCGCACUUAUUC AGAUCCAUUGGCACUCAAGGAAUUCAAGGACUUUGCGGCUGGCA GAAAGUCAAUCGCCCUUGAUCUUGUGACAGAAAUAGGAAGAGU GCCUUCACAUCUAGCCCACAGAACAAGAAACGCUCUGGACAAUC UGGUGAUGCUGCAUACGUCAGAAGAUGGCGGUAGGGCUUACAG GCAUGCGGUGGAGGAACUACCAGAAACAAUGGAAACACUCCUAC UCUUGGGACUAAUGAUCUUGUUGACAGGUGGAGCAAUGCUUUU CUUGAUAUCAGGUAAAGGGAUUGGAAAGACUUCAAUAGGACUC AUUUGUGUAAUCGCUUCCAGCGGCAUGUUGUGGAUGGCCGAAG UUCCACUCCAAUGGAUCGCGUCGGCUAUAGUCCUGGAGUUUUUU AUGAUGGUGUUGCUCAUACCAGAACCAGAAAAGCAGAGAACCCC CCAAGACAACCAACUCGCAUAUGUCGUGAUAGGCAUACUUACAU UGGCUGCAACAAUAGCAGCCAAUGAAAUGGGACUGCUGGAAACC ACAAAGAGAGACUUAGGAAUGUCUAAGGAGCCAGGUGUUGUUU CUCCAACCAGCUAUUUGGAUGUGGACUUGCACCCAGCAUCAGCC UGGACAUUGUACGCCGUGGCCACUACAGUAAUAACACCAAUGUU AAGACAUACCAUAGAGAAUUCUACAGCAAAUGUGUCCCUGGCAG CUAUAGCCAACCAGGCAGUGGUCCUGAUGGGUUUGGACAAAGGA UGGCCAAUAUCAAAAAUGGACUUAGGCGUGCCACUACUGGCACU GGGUUGCUAUUCACAAGUGAACCCACUGACUCUAACUGCGGCAG UACUUUUGCUAAUCACACAUUAUGCUAUCAUAGGUCCAGGAUUG CAAGCAAAAGCCACCCGUGAAGCUCAGAAAAGGACAGCUGCUGG AAUAAUGAAGAAUCCAACAGUGGAUGGGAUAAUGACAAUAGAC CUAGAUUCUGUAAUAUUUGAUUCAAAAUUUGAAAAACAACUGG GACAGGUUAUGCUCCUGGUUUUGUGCGCAGUCCAACUCUUGCUA AUGAGAACAUCAUGGGCCUUGUGUGAAGCUUUAACUCUAGCUAC AGGACCAAUAACAACACUCUGGGAAGGAUCACCUGGUAAGUUCU GGAACACCACGAUAGCUGUUUCCAUGGCGAACAUUUUUAGAGGG AGCUAUUUAGCAGGAGCUGGGCUUGCUUUUUCUAUUAUGAAAU CAGUUGGAACAGGAAAAAGAGGAACAGGCUCACAAGGUGAAAC UUUAGGAGAAAAAUGGAAAAAGAAAUUAAAUCAAUUAUCCCGG AAAGAGUUUGACCUUUACAAGAAAUCUGGAAUCACUGAAGUGG AUAGAACAGAAGCCAAAGAAGGGUUGAAAAGAGGAGAGACAAC ACAUCAUGCCGUGUCCCGAGGUAGCGCAAAACUUCAAUGGUUUG UGGAAAGAAACAUGGUCGUUCCCGAAGGAAGAGUCAUAGACUU GGGCUGUGGAAGAGGAGGCUGGUCAUAUUACUGUGCAGGACUG AAAAAAGUCACAGAAGUGCGAGGAUACACAAAAGGCGGUCCAG GACACGAAGAACCAGUACCUAUGUCUACAUAUGGAUGGAACAUA GUUAAGUUAAUGAGCGGAAAGGAUGUGUUCUAUCUCCCACCUG AAAAGUGUGAUACCCUGUUGUGUGACAUUGGAGAAUCUUCACC AAGCCCAACAGUGGAAGAGAGCAGAACUAUAAGAGUUUUGAAG AUGGUUGAACCAUGGCUAAAAAACAACCAGUUUUGCAUUAAAG UUUUGAACCCUUACAUGCCAACUGUGAUUGAGCACCUAGAAAGA CUACAAAGGAAACAUGGAGGAAUGCUUGUGAGAAAUCCACUUU CACGAAACUCCACGCACGAAAUGUACUGGAUAUCUAAUGGCACA GGUAACAUUGUCUCUUCAGUCAACAUGGUGUCUAGAUUGCUACU GAACAGGUUCACGAUGACACACAGGAGACCCACCAUAGAGAAAG AUGUGGAUUUAGGAGCAGGAACUCGACAUGUUAAUGCGGAACC AGAAACACCCAACAUGGAUGUCAUUGGGGAAAGAAUAAAAAGG AUCAAGGAGGAGCAUAAUUCAACAUGGCACUAUGAUGACGAAA ACCCCUACAAAACGUGGGCUUACCAUGGAUCCUAUGAAGUCAAA GCCACAGGCUCAGCCUCCUCCAUGAUAAAUGGAGUCGUGAAACU CCUCACCAAACCAUGGGAUGUGGUGCCCAUGGUGACACAGAUGG CAAUGACAGACACAACUCCAUUUGGCCAGCAGAGAGUCUUUAAA GAGAAAGUGGACACCAGGACGCCCAGGCCCAUGCCAGGGACAAG AAAGGCUAUGGAGAUCACAGCGGAGUGGCUCUGGAGAACCCUGG GAAGGAACAAAAGACCCAGAUUAUGCACAAGGGAAGAGUUUAC AAAAAAGGUCAGAACUAACGCAGCCAUGGGCGCCGUUUUCACAG AGGAGAACCAAUGGGACAGUGCGAAAGCUGCUGUUGAGGAUGA AGAAUUUUGGAAACUUGUGGACAGAGAACGUGAACUCCACAAA UUGGGCAAAUGUGGAAGCUGCGUUUAUAACAUGAUGGGCAAGA GAGAGAAAAAACUUGGAGAGUUUGGCAAAGCAAAAGGCAGUAG AGCUAUAUGGUACAUGUGGUUGGGAGCCAGGUACCUUGAGUUC GAAGCCCUUGGAUUCUUAAAUGAAGACCACUGGUUCUCGCGUGA AAACUCUUACAGUGGAGUAGAAGGAGAAGGACUGCACAAGCUA GGCUACAUAUUAAGGGACAUUUCCAAGAUACCCGGAGGAGCCAU GUAUGCUGAUGACACAGCUGGUUGGGACACAAGAAUAACAGAA GAUGACCUGCACAAUGAGGAAAAGAUCAUACAGCAAAUGGACCC UGAACACAGGCAGUUAGCGAACGCUAUAUUCAAGCUCACAUACC AAAACAAAGUGGUCAAAGUUCAACGACCGACUCCAACGGGCACG GUAAUGGAUAUUAUAUCUAGGAAAGACCAAAGGGGCAGUGGAC AACUGGGAACUUAUGGCCUGAAUACAUUCACCAACAUGGAAGCC CAGUUAGUCAGACAAAUGGAAGGAGAAGGUGUGCUGACAAAGG CAGACCUCGAGAACCCUCAUCUGCUAGAGAAGAAAAUCACACAA UGGUUGGAAACCAAAGGAGUGGAGAGGUUAAAAAGAAUGGCCA UUAGCGGGGAUGAUUGCGUGGUGAAACCAAUCGAUGACAGGUU CGCUAAUGCCCUGCUUGCUUUGAACGAUAUGGGAAAGGUUCGGA AAGACAUACCUCAAUGGCAGCCAUCAAAGGGAUGGCAUGAUUGG CAACAGGUUCCUUUCUGCUCCCACCACUUUCAUGAAUUGAUCAU GAAAGAUGGAAGAAAGUUGGUGGUUCCCUGCAGACCCCAGGACG AACUAAUAGGAAGAGCAAGAAUCUCUCAAGGAGCGGGAUGGAG CCUUAGAGAAACUGCAUGUCUGGGGAAAGCCUACGCCCAAAUGU GGAGUCUCAUGUAUUUUCACAGAAGAGAUCUCAGAUUAGCAUCC AACGCCAUAUGUUCAGCAGUACCAGUCCACUGGGUUCCCACAAG UAGAACGACAUGGUCUAUUCAUGCUCACCAUCAGUGGAUGACUA CAGAAGACAUGCUUACUGUUUGGAACAGGGUGUGGAUAGAGGA AAAUCCAUGGAUGGAAGACAAAACUCCAGUUACAACUUGGGAA AAUGUUCCAUAUCUAGGAAAGAGAGAAGACCAAUGGUGUGGAU CACUUAUUGGUCUCACUUCCAGAGCAACCUGGGCCCAGAACAUA CCCACAGCAAUUCAACAGGUGAGAAGCCUUAUAGGCAAUGAAGA GUUCCUGGACUACAUGCCUUCAAUGAAGAGAUUCAGGAAGGAA GAGGAGUCGGAGGGAGCCAUUUGGUAAACGUAGGAAGUGGAAA AGAGGCUAACUGUCAGGCCACCUUAAGCCACAGUACGGAAGAAG CUGUGCUGCCUGUGAGCCCCGUCCAAGGACGUUAAAAGAAGAAG UCAGGCCCCAAAGCCACGGUUUGAGCAAACCGUGCUGCCUGUAG CUCCGUCGUGGGGACGUAAAACCUGGGAGGCUGCAAACUGUGGA AGCUGUACGCACGGUGUAGCAGACUAGCGGUUAGAGGAGACCCC UCCCAUGACACAACGCAGCAGCGGGGCCCGAGCACUGAGGGAAG CUGUACCUCCUUGCAAAGGACUAGAGGUUAGAGGAGACCCCCCG CAAAUAAAAACAGCAUAUUGACGCUGGGAGAGACCAGAGAUCCU GCUGUCUCCUCAGCAUCAUUCCAGGCACAGAACGCCAGAAAAUG GAAUGGUGCUGUUGAAUCAACAGGUUCU DEN-4 AGUUGUUAGUCUGUGUGGACCGACAAGGACAGUUCCAAAUCGG 243 (NC_002640.1) AAGCUUGCUUAACACAGUUCUAACAGUUUGUUUGAAUAGAGAG CAGAUCUCUGGAAAAAUGAACCAACGAAAAAAGGUGGUUAGAC CACCUUUCAAUAUGCUGAAACGCGAGAGAAACCGCGUAUCAACC CCUCAAGGGUUGGUGAAGAGAUUCUCAACCGGACUUUUUUCUGG GAAAGGACCCUUACGGAUGGUGCUAGCAUUCAUCACGUUUUUGC GAGUCCUUUCCAUCCCACCAACAGCAGGGAUUCUGAAGAGAUGG GGACAGUUGAAGAAAAAUAAGGCCAUCAAGAUACUGAUUGGAU UCAGGAAGGAGAUAGGCCGCAUGCUGAACAUCUUGAACGGGAG AAAAAGGUCAACGAUAACAUUGCUGUGCUUGAUUCCCACCGUAA UGGCGUUUUCCCUCAGCACAAGAGAUGGCGAACCCCUCAUGAUA GUGGCAAAACAUGAAAGGGGGAGACCUCUCUUGUUUAAGACAA CAGAGGGGAUCAACAAAUGCACUCUCAUUGCCAUGGACUUGGGU GAAAUGUGUGAGGACACUGUCACGUAUAAAUGCCCCCUACUGGU CAAUACCGAACCUGAAGACAUUGAUUGCUGGUGCAACCUCACGU CUACCUGGGUCAUGUAUGGGACAUGCACCCAGAGCGGAGAACGG AGACGAGAGAAGCGCUCAGUAGCUUUAACACCACAUUCAGGAAU GGGAUUGGAAACAAGAGCUGAGACAUGGAUGUCAUCGGAAGGG GCUUGGAAGCAUGCUCAGAGAGUAGAGAGCUGGAUACUCAGAA ACCCAGGAUUCGCGCUCUUGGCAGGAUUUAUGGCUUAUAUGAUU GGGCAAACAGGAAUCCAGCGAACUGUCUUCUUUGUCCUAAUGAU GCUGGUCGCCCCAUCCUACGGAAUGCGAUGCGUAGGAGUAGGAA ACAGAGACUUUGUGGAAGGAGUCUCAGGUGGAGCAUGGGUCGA CCUGGUGCUAGAACAUGGAGGAUGCGUCACAACCAUGGCCCAGG GAAAACCAACCUUGGAUUUUGAACUGACUAAGACAACAGCCAAG GAAGUGGCUCUGUUAAGAACCUAUUGCAUUGAAGCCUCAAUAUC AAACAUAACUACGGCAACAAGAUGUCCAACGCAAGGAGAGCCUU AUCUGAAAGAGGAACAGGACCAACAGUACAUUUGCCGGAGAGA UGUGGUAGACAGAGGGUGGGGCAAUGGCUGUGGCUUGUUUGGA AAAGGAGGAGUUGUGACAUGUGCGAAGUUUUCAUGUUCGGGGA AGAUAACAGGCAAUUUGGUCCAAAUUGAGAACCUUGAAUACAC AGUGGUUGUAACAGUCCACAAUGGAGACACCCAUGCAGUAGGAA AUGACACAUCCAAUCAUGGAGUUACAGCCAUGAUAACUCCCAGG UCACCAUCGGUGGAAGUCAAAUUGCCGGACUAUGGAGAACUAAC ACUCGAUUGUGAACCCAGGUCUGGAAUUGACUUUAAUGAGAUG AUUCUGAUGAAAAUGAAAAAGAAAACAUGGCUCGUGCAUAAGC AAUGGUUUUUGGAUCUGCCUCUUCCAUGGACAGCAGGAGCAGAC ACAUCAGAGGUUCACUGGAAUUACAAAGAGAGAAUGGUGACAU UUAAGGUUCCUCAUGCCAAGAGACAGGAUGUGACAGUGCUGGG AUCUCAGGAAGGAGCCAUGCAUUCUGCCCUCGCUGGAGCCACAG AAGUGGACUCCGGUGAUGGAAAUCACAUGUUUGCAGGACAUCU UAAGUGCAAAGUCCGUAUGGAGAAAUUGAGAAUCAAGGGAAUG UCAUACACGAUGUGUUCAGGAAAGUUUUCAAUUGACAAAGAGA UGGCAGAAACACAGCAUGGGACAACAGUGGUGAAAGUCAAGUA UGAAGGUGCUGGAGCUCCGUGUAAAGUCCCCAUAGAGAUAAGA GAUGUAAACAAGGAAAAAGUGGUUGGGCGUAUCAUCUCAUCCA CCCCUUUGGCUGAGAAUACCAACAGUGUAACCAACAUAGAAUUA GAACCCCCCUUUGGGGACAGCUACAUAGUGAUAGGUGUUGGAAA CAGCGCAUUAACACUCCAUUGGUUCAGGAAAGGGAGUUCCAUUG GCAAGAUGUUUGAGUCCACAUACAGAGGUGCAAAACGAAUGGCC AUUCUAGGUGAAACAGCUUGGGAUUUUGGUUCCGUUGGUGGAC UGUUCACAUCAUUGGGAAAGGCUGUGCACCAGGUUUUUGGAAG UGUGUAUACAACCAUGUUUGGAGGAGUCUCAUGGAUGAUUAGA AUCCUAAUUGGGUUCUUAGUGUUGUGGAUUGGCACGAACUCGA GGAACACUUCAAUGGCUAUGACGUGCAUAGCUGUUGGAGGAAU CACUCUGUUUCUGGGCUUCACAGUUCAAGCAGACAUGGGUUGUG UGGCGUCAUGGAGUGGGAAAGAAUUGAAGUGUGGAAGCGGAAU UUUUGUGGUUGACAACGUGCACACUUGGACAGAACAGUACAAA UUUCAACCAGAGUCCCCAGCGAGACUAGCGUCUGCAAUAUUAAA UGCCCACAAAGAUGGGGUCUGUGGAAUUAGAUCAACCACGAGGC UGGAAAAUGUCAUGUGGAAGCAAAUAACCAACGAGCUAAACUA UGUUCUCUGGGAAGGAGGACAUGACCUCACUGUAGUGGCUGGG GAUGUGAAGGGGGUGUUGACCAAAGGCAAGAGAGCACUCACACC CCCAGUGAGUGAUCUGAAAUAUUCAUGGAAGACAUGGGGAAAA GCAAAAAUCUUCACCCCAGAAGCAAGAAAUAGCACAUUUUUAAU AGACGGACCAGACACCUCUGAAUGCCCCAAUGAACGAAGAGCAU GGAACUCUCUUGAGGUGGAAGACUAUGGAUUUGGCAUGUUCAC GACCAACAUAUGGAUGAAAUUCCGAGAAGGAAGUUCAGAAGUG UGUGACCACAGGUUAAUGUCAGCUGCAAUUAAAGAUCAGAAAG CUGUGCAUGCUGACAUGGGUUAUUGGAUAGAGAGCUCAAAAAA CCAGACCUGGCAGAUAGAGAAAGCAUCUCUUAUUGAAGUGAAA ACAUGUCUGUGGCCCAAGACCCACACACUGUGGAGCAAUGGAGU GCUGGAAAGCCAGAUGCUCAUUCCAAAAUCAUAUGCGGGCCCUU UUUCACAGCACAAUUACCGCCAGGGCUAUGCCACGCAAACCGUG GGCCCAUGGCACUUAGGCAAAUUAGAGAUAGACUUUGGAGAAU GCCCCGGAACAACAGUCACAAUUCAGGAGGAUUGUGACCAUAGA GGCCCAUCUUUGAGGACCACCACUGCAUCUGGAAAACUAGUCAC GCAAUGGUGCUGCCGCUCCUGCACGAUGCCUCCCUUAAGGUUCU UGGGAGAAGAUGGGUGCUGGUAUGGGAUGGAGAUUAGGCCCUU GAGUGAAAAAGAAGAGAACAUGGUCAAAUCACAGGUGACGGCC GGACAGGGCACAUCAGAAACUUUUUCUAUGGGUCUGUUGUGCCU GACCUUGUUUGUGGAAGAAUGCUUGAGGAGAAGAGUCACUAGG AAACACAUGAUAUUAGUUGUGGUGAUCACUCUUUGUGCUAUCA UCCUGGGAGGCCUCACAUGGAUGGACUUACUACGAGCCCUCAUC AUGUUGGGGGACACUAUGUCUGGUAGAAUAGGAGGACAGAUCC ACCUAGCCAUCAUGGCAGUGUUCAAGAUGUCACCAGGAUACGUG CUGGGUGUGUUUUUAAGGAAACUCACUUCAAGAGAGACAGCAC UAAUGGUAAUAGGAAUGGCCAUGACAACGGUGCUUUCAAUUCC ACAUGACCUUAUGGAACUCAUUGAUGGAAUAUCACUGGGACUA AUUUUGCUAAAAAUAGUAACACAGUUUGACAACACCCAAGUGG GAACCUUAGCUCUUUCCUUGACUUUCAUAAGAUCAACAAUGCCA UUGGUCAUGGCUUGGAGGACCAUUAUGGCUGUGUUGUUUGUGG UCACACUCAUUCCUUUGUGCAGGACAAGCUGUCUUCAAAAACAG UCUCAUUGGGUAGAAAUAACAGCACUCAUCCUAGGAGCCCAAGC UCUGCCAGUGUACCUAAUGACUCUUAUGAAAGGAGCCUCAAGAA GAUCUUGGCCUCUUAACGAGGGCAUAAUGGCUGUGGGUUUGGU UAGUCUCUUAGGAAGCGCUCUUUUAAAGAAUGAUGUCCCUUUA GCUGGCCCAAUGGUGGCAGGAGGCUUACUUCUGGCGGCUUACGU GAUGAGUGGUAGCUCAGCAGAUCUGUCACUAGAGAAGGCCGCCA ACGUGCAGUGGGAUGAAAUGGCAGACAUAACAGGCUCAAGCCCA AUCGUAGAAGUGAAGCAGGAUGAAGAUGGCUCUUUCUCCAUAC GGGACGUCGAGGAAACCAAUAUGAUAACCCUUUUGGUGAAACU GGCACUGAUAACAGUGUCAGGUCUCUACCCCUUGGCAAUUCCAG UCACAAUGACCUUAUGGUACAUGUGGCAAGUGAAAACACAAAG AUCAGGAGCCCUGUGGGACGUCCCCUCACCCGCUGCCACUAAAA AAGCCGCACUGUCUGAAGGAGUGUACAGGAUCAUGCAAAGAGG GUUAUUCGGGAAAACUCAGGUUGGAGUAGGGAUACACAUGGAA GGUGUAUUUCACACAAUGUGGCAUGUAACAAGAGGAUCAGUGA UCUGCCACGAGACUGGGAGAUUGGAGCCAUCUUGGGCUGACGUC AGGAAUGACAUGAUAUCAUACGGUGGGGGAUGGAGGCUUGGAG ACAAAUGGGACAAAGAAGAAGACGUUCAGGUCCUCGCCAUAGAA CCAGGAAAAAAUCCUAAACAUGUCCAAACGAAACCUGGCCUUUU CAAGACCCUAACUGGAGAAAUUGGAGCAGUAACAUUAGAUUUC AAACCCGGAACGUCUGGUUCUCCCAUCAUCAACAGGAAAGGAAA AGUCAUCGGACUCUAUGGAAAUGGAGUAGUUACCAAAUCAGGU GAUUACGUCAGUGCCAUAACGCAAGCCGAAAGAAUUGGAGAGCC AGAUUAUGAAGUGGAUGAGGACAUUUUUCGAAAGAAAAGAUUA ACUAUAAUGGACUUACACCCCGGAGCUGGAAAGACAAAAAGAAU UCUUCCAUCAAUAGUGAGAGAAGCCUUAAAAAGGAGGCUACGA ACUUUGAUUUUAGCUCCCACGAGAGUGGUGGCGGCCGAGAUGGA AGAGGCCCUACGUGGACUGCCAAUCCGUUAUCAGACCCCAGCUG UGAAAUCAGAACACACAGGAAGAGAGAUUGUAGACCUCAUGUG UCAUGCAACCUUCACAACAAGACUUUUGUCAUCAACCAGGGUUC CAAAUUACAACCUUAUAGUGAUGGAUGAAGCACAUUUCACCGAU CCUUCUAGUGUCGCGGCUAGAGGAUACAUCUCGACCAGGGUGGA AAUGGGAGAGGCAGCAGCCAUCUUCAUGACCGCAACCCCUCCCG GAGCGACAGAUCCCUUUCCCCAGAGCAACAGCCCAAUAGAAGAC AUCGAGAGGGAAAUUCCGGAAAGGUCAUGGAACACAGGGUUCG ACUGGAUAACAGACUACCAAGGGAAAACUGUGUGGUUUGUUCCC AGCAUAAAAGCUGGAAAUGACAUUGCAAAUUGUUUGAGAAAGU CGGGAAAGAAAGUUAUCCAGUUGAGUAGGAAAACCUUUGAUAC AGAGUAUCCAAAAACGAAACUCACGGACUGGGACUUUGUGGUCA CUACAGACAUAUCUGAAAUGGGGGCCAAUUUUAGAGCCGGGAG AGUGAUAGACCCUAGAAGAUGCCUCAAGCCAGUUAUCCUACCAG AUGGGCCAGAGAGAGUCAUUUUAGCAGGUCCUAUUCCAGUGACU CCAGCAAGCGCUGCUCAGAGAAGAGGGCGAAUAGGAAGGAACCC AGCACAAGAAGACGACCAAUACGUUUUCUCCGGAGACCCACUAA AAAAUGAUGAAGAUCAUGCCCACUGGACAGAAGCAAAGAUGCU GCUUGACAAUAUCUACACCCCAGAAGGGAUCAUUCCAACAUUGU UUGGUCCGGAAAGGGAAAAAACCCAAGCCAUUGAUGGAGAGUU UCGCCUCAGAGGGGAACAAAGGAAGACUUUUGUGGAAUUAAUG AGGAGAGGAGACCUUCCGGUGUGGCUGAGCUAUAAGGUAGCUU CUGCUGGCAUUUCUUACGAAGAUCGGGAAUGGUGCUUCACAGGG GAAAGAAAUAACCAAAUUUUAGAAGAAAACAUGGAGGUUGAAA UUUGGACUAGAGAGGGAGAAAAGAAAAAGCUAAGGCCAAGAUG GUUAGAUGCACGUGUAUACGCUGACCCCAUGGCUUUGAAGGAUU UCAAGGAGUUUGCCAGUGGAAGGAAGAGUAUAACUCUCGACAU CCUAACAGAGAUUGCCAGUUUGCCAACUUACCUUUCCUCUAGGG CCAAGCUCGCCCUUGAUAACAUAGUCAUGCUCCACACAACAGAA AGAGGAGGGAGGGCCUAUCAACACGCCCUGAACGAACUUCCGGA GUCACUGGAAACACUCAUGCUUGUAGCUUUACUAGGUGCUAUGA CAGCAGGCAUCUUCCUGUUUUUCAUGCAAGGGAAAGGAAUAGG GAAAUUGUCAAUGGGUUUGAUAACCAUUGCGGUGGCUAGUGGC UUGCUCUGGGUAGCAGAAAUUCAACCCCAGUGGAUAGCGGCCUC AAUCAUACUAGAGUUUUUUCUCAUGGUACUGUUGAUACCGGAA CCAGAAAAACAAAGGACCCCACAAGACAAUCAAUUGAUCUACGU CAUAUUGACCAUUCUCACCAUCAUUGGUCUAAUAGCAGCCAACG AGAUGGGGCUGAUUGAAAAAACAAAAACGGAUUUUGGGUUUUA CCAGGUAAAAACAGAAACCACCAUCCUCGAUGUGGACUUGAGAC CAGCUUCAGCAUGGACGCUCUAUGCAGUAGCCACCACAAUUCUG ACUCCCAUGCUGAGACACACCAUAGAAAACACGUCGGCCAACCU AUCUCUAGCAGCCAUUGCCAACCAGGCAGCCGUCCUAAUGGGGC UUGGAAAAGGAUGGCCGCUCCACAGAAUGGACCUCGGUGUGCCG CUGUUAGCAAUGGGAUGCUAUUCUCAAGUGAACCCAACAACCUU GACAGCAUCCUUAGUCAUGCUUUUAGUCCAUUAUGCAAUAAUAG GCCCAGGAUUGCAGGCAAAAGCCACAAGAGAGGCCCAGAAAAGG ACAGCUGCUGGGAUCAUGAAAAAUCCCACAGUGGACGGGAUAAC AGUAAUAGAUCUAGAACCAAUAUCCUAUGACCCAAAAUUUGAA AAGCAAUUAGGGCAGGUCAUGCUACUAGUCUUGUGUGCUGGAC AACUACUCUUGAUGAGAACAACAUGGGCUUUCUGUGAAGUCUU GACUUUGGCCACAGGACCAAUCUUGACCUUGUGGGAGGGCAACC CGGGAAGGUUUUGGAACACGACCAUAGCCGUAUCCACCGCCAAC AUUUUCAGGGGAAGUUACUUGGCGGGAGCUGGACUGGCUUUUU CACUCAUAAAGAAUGCACAAACCCCUAGGAGGGGAACUGGGACC ACAGGAGAGACACUGGGAGAGAAGUGGAAGAGACAGCUAAACU CAUUAGACAGAAAAGAGUUUGAAGAGUAUAAAAGAAGUGGAAU ACUAGAAGUGGACAGGACUGAAGCCAAGUCUGCCCUGAAAGAUG GGUCUAAAAUCAAGCAUGCAGUAUCAAGAGGGUCCAGUAAGAU CAGAUGGAUUGUUGAGAGAGGGAUGGUAAAGCCAAAAGGGAAA GUUGUAGAUCUUGGCUGUGGGAGAGGAGGAUGGUCUUAUUACA UGGCGACACUCAAGAACGUGACUGAAGUGAAAGGGUAUACAAA AGGAGGUCCAGGACAUGAAGAACCGAUUCCCAUGGCUACUUAUG GUUGGAAUUUGGUCAAACUCCAUUCAGGGGUUGACGUGUUCUA CAAACCCACAGAGCAAGUGGACACCCUGCUCUGUGAUAUUGGGG AGUCAUCUUCUAAUCCAACAAUAGAGGAAGGAAGAACAUUAAG AGUUUUGAAGAUGGUGGAGCCAUGGCUCUCUUCAAAACCUGAA UUCUGCAUCAAAGUCCUUAACCCCUACAUGCCAACAGUCAUAGA AGAGCUGGAGAAACUGCAGAGAAAACAUGGUGGGAACCUUGUC AGAUGCCCGCUGUCCAGGAACUCCACCCAUGAGAUGUAUUGGGU GUCAGGAGCGUCGGGAAACAUUGUGAGCUCUGUGAACACAACAU CAAAGAUGUUGUUGAACAGGUUCACAACAAGGCAUAGGAAACCC ACUUAUGAGAAGGACGUAGAUCUUGGGGCAGGAACGAGAAGUG UCUCCACUGAAACAGAAAAACCAGACAUGACAAUCAUUGGGAGA AGGCUUCAGCGAUUGCAAGAAGAGCACAAAGAAACCUGGCAUUA UGAUCAGGAAAACCCAUACAGAACCUGGGCGUAUCAUGGAAGCU AUGAAGCUCCUUCGACAGGCUCUGCAUCCUCCAUGGUGAACGGG GUGGUAAAACUGCUAACAAAACCCUGGGAUGUGAUUCCAAUGG UGACUCAGUUAGCCAUGACAGAUACAACCCCUUUUGGGCAACAA AGAGUGUUCAAAGAGAAGGUGGAUACCAGAACACCACAACCAAA ACCCGGUACACGAAUGGUUAUGACCACGACAGCCAAUUGGCUGU GGGCCCUCCUUGGAAAGAAGAAAAAUCCCAGACUGUGCACAAGG GAAGAGUUCAUCUCAAAAGUUAGAUCAAACGCAGCCAUAGGCGC AGUCUUUCAGGAAGAACAGGGAUGGACAUCAGCCAGUGAAGCU GUGAAUGACAGCCGGUUUUGGGAACUGGUUGACAAAGAAAGGG CCCUACACCAGGAAGGGAAAUGUGAAUCGUGUGUCUAUAACAUG AUGGGAAAACGUGAGAAAAAGUUAGGAGAGUUUGGCAGAGCCA AGGGAAGCCGAGCAAUCUGGUACAUGUGGCUGGGAGCGCGGUU UCUGGAAUUUGAAGCCCUGGGUUUUUUGAAUGAAGAUCACUGG UUUGGCAGAGAAAAUUCAUGGAGUGGAGUGGAAGGGGAAGGUC UGCACAGAUUGGGAUAUAUCCUGGAGGAGAUAGACAAGAAGGA UGGAGACCUAAUGUAUGCUGAUGACACAGCAGGCUGGGACACAA GAAUCACUGAGGAUGACCUUCAAAAUGAGGAACUGAUCACGGA ACAGAUGGCUCCCCACCACAAGAUCCUAGCCAAAGCCAUUUUCA AACUAACCUAUCAAAACAAAGUGGUGAAAGUCCUCAGACCCACA CCGCGGGGAGCGGUGAUGGAUAUCAUAUCCAGGAAAGACCAAAG AGGUAGUGGACAAGUUGGAACAUAUGGUUUGAACACAUUCACC AACAUGGAAGUUCAACUCAUCCGCCAAAUGGAAGCUGAAGGAGU CAUCACACAAGAUGACAUGCAGAACCCAAAAGGGUUGAAAGAAA GAGUUGAGAAAUGGCUGAAAGAGUGUGGUGUCGACAGGUUAAA GAGGAUGGCAAUCAGUGGAGACGAUUGCGUGGUGAAGCCCCUA GAUGAGAGGUUUGGCACUUCCCUCCUCUUCUUGAACGACAUGGG AAAGGUGAGGAAAGACAUUCCGCAGUGGGAACCAUCUAAGGGA UGGAAAAACUGGCAAGAGGUUCCUUUUUGCUCCCACCACUUUCA CAAGAUCUUUAUGAAGGAUGGCCGCUCACUAGUUGUUCCAUGUA GAAACCAGGAUGAACUGAUAGGGAGAGCCAGAAUCUCGCAGGG AGCUGGAUGGAGCUUAAGAGAAACAGCCUGCCUGGGCAAAGCUU ACGCCCAGAUGUGGUCGCUUAUGUACUUCCACAGAAGGGAUCUG CGUUUAGCCUCCAUGGCCAUAUGCUCAGCAGUUCCAACGGAAUG GUUUCCAACAAGCAGAACAACAUGGUCAAUCCACGCUCAUCACC AGUGGAUGACCACUGAAGAUAUGCUCAAAGUGUGGAACAGAGU GUGGAUAGAAGACAACCCUAAUAUGACUGACAAGACUCCAGUCC AUUCGUGGGAAGAUAUACCUUACCUAGGGAAAAGAGAGGAUUU GUGGUGUGGAUCCCUGAUUGGACUUUCUUCCAGAGCCACCUGGG CGAAGAACAUUCAUACGGCCAUAACCCAGGUCAGGAACCUGAUC GGAAAAGAGGAAUACGUGGAUUACAUGCCAGUAAUGAAAAGAU ACAGUGCUCCUUCAGAGAGUGAAGGAGUUCUGUAAUUACCAACA ACAAACACCAAAGGCUAUUGAAGUCAGGCCACUUGUGCCACGGU UUGAGCAAACCGUGCUGCCUGUAGCUCCGCCAAUAAUGGGAGGC GUAAUAAUCCCCAGGGAGGCCAUGCGCCACGGAAGCUGUACGCG UGGCAUAUUGGACUAGCGGUUAGAGGAGACCCCUCCCAUCACUG AUAAAACGCAGCAAAAGGGGGCCCGAAGCCAGGAGGAAGCUGUA CUCCUGGUGGAAGGACUAGAGGUUAGAGGAGACCCCCCCAACAC AAAAACAGCAUAUUGACGCUGGGAAAGACCAGAGAUCCUGCUGU CUCUGCAACAUCAAUCCAGGCACAGAGCGCCGCAAGAUGGAUUG GUGUUGUUGAUCCAACAGGUUCU Construct 1 AUGGAUGCUAUGAAAAGAGGCCUGUGUUGUGUGUUGCUGUUGU 244 GCGGAGCUGUGUUUGUGUCACCUUUCCACCUGACUACCCGCAAU GGUGAGCCCCAUAUGAUUGUGUCGCGCCAGGAGAAGGGGAAGUC CCUCCUGUUCAAAACUGAAAACGGCGUGAACAUGUGUACCCUGA UGGCCAUGGACCUUGGAGAACUGUGCGAGGACACCAUCACCUAC AAUUGUCCGCUCCUGCGCCAAAACGAACCAGAAGAUAUCGACUG CUGGUGCAAUUCCACUUCAACCUGGGUUACCUACGGAACUUGCA CCGCCACGGGAGAACACAGAAGAGAAAAGCGCUCGGUGGCGCUG GUGCCUCAUGUCGGAAUGGGACUGGAGACUCGGACGGAGACUUG GAUGUCCUCGGAGGGAGCAUGGAAACAUGCCCAACGGAUCGAAA CUUGGGUGCUGAGGCACCCUGGAUUCACCAUCAUGGCAGCGAUC CUCGCCUACACUAUAGGUACUACCUACUUUCAAAGGGUGCUGAU CUUCAUUCUCCUCACCGCAGUGGCCCCUUCAAUGACCAUGAGGU GCAUUGGGAUCUCGAACCGGGACUUCGUCGAAGGAGUGUCCGGA GGUAGCUGGGUCGACAUCGUCCUGGAACACGGAAGCUGCGUGAC UACUAUGGCGAAGAACAAGCCAACCUUGGACUUCGAGCUUAUCA AGACCGAGGCGAAGCACCCGGCCACUCUGAGAAAGUACUGCAUC GAGGCUAAGCUCACCAACACGACCACUGCCUCGCGAUGCCCAAC UCAGGGAGAACCGUCACUGAACGAAGAACAGGAUAAACGCUUUG UGUGCAAGCAUAGCAUGGUGGAUAGAGGCUGGGGAAACGGCUG UGGACUCUUCGGAAAGGGUGGAAUUGUGACGUGCGCAAUGUUC ACUUGCAAGAAGAAUAUGGAAGGGAAGAUCGUCCAGCCGGAGA ACCUGGAAUACACUAUCGUGAUCACCCCGCACUCAGGCGAGGAG AACGCAGUGGGCAACGAUACCGGGAAGCACGGGAAGGAAAUCAA GGUGACCCCGCAGUCGUCCAUUACCGAGGCCGAACUCACCGGAU ACGGCACUGUGACUAUGGAAUGCUCGCCACGGACCGGGCUGGAU UUCAAUGAGAUGGUGCUCUUGCAAAUGGAGAACAAAGCCUGGC UGGUCCACCGCCAGUGGUUCCUCGACCUCCCCCUUCCGUGGCUG CCGGGAGCUGACACCCAAGGAUCCAACUGGAUCCAAAAAGAAAC CCUUGUCACGUUUAAGAAUCCACAUGCCAAAAAGCAGGACGUGG UCGUGCUCGGAAGCCAGGAAGGAGCCAUGCACACUGCGCUGACU GGAGCAACCGAAAUUCAAAUGUCGAGCGGCAACCUCCUCUUCAC UGGACAUCUGAAGUGCCGGCUGCGCAUGGACAAACUGCAACUUA AGGGCAUGUCAUACUCGAUGUGUACCGGCAAAUUCAAGGUGGU GAAGGAGAUCGCGGAGACUCAGCACGGGACCAUCGUCAUCCGGG UCCAGUAUGAGGGUGAUGGUUCCCCCUGCAAGAUCCCUUUCGAA AUCAUGGAUCUGGAGAAACGUCACGUGCUGGGCCGGCUGAUCAC UGUGAAUCCGAUCGUUACGGAGAAAGACAGCCCGGUGAACAUCG AAGCUGAACCGCCGUUUGGGGAUAGCUACAUUAUCAUCGGCGUG GAACCAGGCCAGCUCAAGUUGUCGUGGUUCAAGAAAGGAUCCAG CAUCGGACAGAUGUUCGAAACCACUAUGCGCGGAGCCAAACGCA UGGCUAUCCUGGGGGACACGGCCUGGGACUUCGGGUCGCUGGGU GGUGUGUUCACCUCCAUUGGAAAGGCGCUCCAUCAGGUGUUUGG UGCGAUCUACGGCGCCGCAUUCUCCGGAGUGUCAUGGACCAUGA AGAUCCUCAUCGGAGUCGUCAUCACCUGGAUCGGCAUGAAUUCU CGGUCCACUUCCUUGAGCGUCAGCCUGGUGCUGGUCGGAGUUGU GACUCUGUACCUUGGAGUGAUGGUCCAGGCC Construct 2 AUGGAUGCUAUGAAAAGAGGCCUGUGUUGUGUGUUGCUGUUGU 245 GCGGAGCUGUGUUUGUGUCACCUUUCCACCUGACUACCCGCAAU GGUGAGCCCCAUAUGAUUGUGUCGCGCCAGGAGAAGGGGAAGUC CCUCCUGUUCAAAACUGAAAACGGCGUGAACAUGUGUACCCUGA UGGCCAUGGACCUUGGAGAACUGUGCGAGGACACCAUCACCUAC AAUUGUCCGCUCCUGCGCCAAAACGAACCAGAAGAUAUCGACUG CUGGUGCAAUUCCACUUCAACCUGGGUUACCUACGGAACUUGCA CCGCCACGGGAGAACACAGAAGAGAAAAGCGCUCGGUGGCGCUG GUGCCUCAUGUCGGAAUGGGACUGGAGACUCGGACGGAGACUUG GAUGUCCUCGGAGGGAGCAUGGAAACAUGCCCAACGGAUCGAAA CUUGGGUGCUGAGGCACCCUGGAUUCACCAUCAUGGCAGCGAUC CUCGCCUACACUAUAGGUACUACCUACUUUCAAAGGGUGCUGAU CUUCAUUCUCCUCACCGCAGUGGCCCCUUCAAUGACCAUGAGGU GCAUUGGGAUCUCGAACCGGGACUUCGUCGAAGGAGUGUCCGGA GGUAGCUGGGUCGACAUCGUCCUGGAACACGGAAGCUGCGUGAC UACUAUGGCGAAGAACAAGCCAACCUUGGACUUCGAGCUUAUCA AGACCGAGGCGAAGCACCCGGCCACUCUGAGAAAGUACUGCAUC GAGGCUAAGCUCACCAACACGACCACUGCCUCGCGAUGCCCAAC UCAGGGAGAACCGUCACUGAACGAAGAACAGGAUAAACGCUUUG UGUGCAAGCAUAGCAUGGUGGAUAGAGGCUGGGGAAACGGCUG UGGACUCUUCGGAAAGGGUGGAAUUGUGACGUGCGCAAUGUUC ACUUGCAAGAAGAAUAUGGAAGGGAAGAUCGUCCAGCCGGAGA ACCUGGAAUACACUAUCGUGAUCACCCCGCACUCAGGCGAGGAG AACGCAGUGGGCAACGAUACCGGGAAGCACGGGAAGGAAAUCAA GGUGACCCCGCAGUCGUCCAUUACCGAGGCCGAACUCACCGGAU ACGGCACUGUGACUAUGGAAUGCUCGCCACGGACCGGGCUGGAU UUCAAUGAGAUGGUGCUCUUGCAAAUGGAGAACAAAGCCUGGC UGGUCCACCGCCAGUGGUUCCUCGACCUCCCCCUUCCGUGGCUG CCGGGAGCUGACACCCAAGGAUCCAACUGGAUCCAAAAAGAAAC CCUUGUCACGUUUAAGAAUCCACAUGCCAAAAAGCAGGACGUGG UCGUGCUCGGAAGCCAGGAAGGAGCCAUGCACACUGCGCUGACU GGAGCAACCGAAAUUCAAAUGUCGAGCGGCAACCUCCUCUUCAC UGGACAUCUGAAGUGCCGGCUGCGCAUGGACAAACUGCAACUUA AGGGCAUGUCAUACUCGAUGUGUACCGGCAAAUUCAAGGUGGU GAAGGAGAUCGCGGAGACUCAGCACGGGACCAUCGUCAUCCGGG UCCAGUAUGAGGGUGAUGGUUCCCCCUGCAAGAUCCCUUUCGAA AUCAUGGAUCUGGAGAAACGUCACGUGCUGGGCCGGCUGAUCAC UGUGAAUCCGAUCGUUACGGAGAAAGACAGCCCGGUGAACAUCG AAGCUGAACCGCCGUUUGGGGAUAGCUACAUUAUCAUCGGCGUG GAACCAGGCCAGCUCAAGUUGUCGUGGUUCAAGAAAGGA Construct 3 AUGGAUGCUAUGAAAAGAGGCCUGUGUUGUGUGUUGCUGUUGU 246 GCGGAGCUGUGUUUGUGUCACCUUUCCACCUGACUACCCGCAAU GGUGAGCCCCAUAUGAUUGUGUCGCGCCAGGAGAAGGGGAAGUC CCUCCUGUUCAAAACUGAAAACGGCGUGAACAUGUGUACCCUGA UGGCCAUGGACCUUGGAGAACUGUGCGAGGACACCAUCACCUAC AAUUGUCCGCUCCUGCGCCAAAACGAACCAGAAGAUAUCGACUG CUGGUGCAAUUCCACUUCAACCUGGGUUACCUACGGAACUUGCA CCGCCACGGGAGAACACAGAAGAGAAAAGCGCUCGGUGGCGCUG GUGCCUCAUGUCGGAAUGGGACUGGAGACUCGGACGGAGACUUG GAUGUCCUCGGAGGGAGCAUGGAAACAUGCCCAACGGAUCGAAA CUUGGGUGCUGAGGCACCCUGGAUUCACCAUCAUGGCAGCGAUC CUCGCCUACACUAUAGGUACUACCUACUUUCAAAGGGUGCUGAU CUUCAUUCUCCUCACCGCAGUGGCCCCUUCAAUGACCAUGAGGU GCAUUGGGAUCUCGAACCGGGACUUCGUCGAAGGAGUGUCCGGA GGUAGCUGGGUCGACAUCGUCCUGGAACACGGAAGCUGCGUGAC UACUAUGGCGAAGAACAAGCCAACCUUGGACUUCGAGCUUAUCA AGACCGAGGCGAAGCACCCGGCCACUCUGAGAAAGUACUGCAUC GAGGCUAAGCUCACCAACACGACCACUGCCUCGCGAUGCCCAAC UCAGGGAGAACCGUCACUGAACGAAGAACAGGAUAAACGCUUUG UGUGCAAGCAUAGCAUGGUGGAUAGAGGCUGGGGAAACGGCUG UGGACUCUUCGGAAAGGGUGGAAUUGUGACGUGCGCAAUGUUC ACUUGCAAGAAGAAUAUGGAAGGGAAGAUCGUCCAGCCGGAGA ACCUGGAAUACACUAUCGUGAUCACCCCGCACUCAGGCGAGGAG AACGCAGUGGGCAACGAUACCGGGAAGCACGGGAAGGAAAUCAA GGUGACCCCGCAGUCGUCCAUUACCGAGGCCGAACUCACCGGAU ACGGCACUGUGACUAUGGAAUGCUCGCCACGGACCGGGCUGGAU UUCAAUGAGAUGGUGCUCUUGCAAAUGGAGAACAAAGCCUGGC UGGUCCACCGCCAGUGGUUCCUCGACCUCCCCCUUCCGUGGCUG CCGGGAGCUGACACCCAAGGAUCCAACUGGAUCCAAAAAGAAAC CCUUGUCACGUUUAAGAAUCCACAUGCCAAAAAGCAGGACGUGG UCGUGCUCGGAAGCCAGGAAGGAGCCAUGCACACUGCGCUGACU GGAGCAACCGAAAUUCAAAUGUCGAGCGGCAACCUCCUCUUCAC UGGACAUCUGAAGUGCCGGCUGCGCAUGGACAAACUGCAACUUA AGGGCAUGUCAUACUCGAUGUGUACCGGCAAAUUCAAGGUGGU GAAGGAGAUCGCGGAGACUCAGCACGGGACCAUCGUCAUCCGGG UCCAGUAUGAGGGUGAUGGUUCCCCCUGCAAGAUCCCUUUCGAA AUCAUGGAUCUGGAGAAACGUCACGUGCUGGGCCGGCUGAUCAC UGUGAAUCCGAUCGUUACGGAGAAAGACAGCCCGGUGAACAUCG AAGCUGAACCGCCGUUUGGGGAUAGCUACAUUAUCAUCGGCGUG GAACCAGGCCAGCUCAAGUUGUCGUGGUUCAAGAAAGGAGGAG GUGGAGGAUCCGGAGGCGGAGGGUCGGGCGGUGGUGGAUCGGA GGUCAAACUGCAGCAAUCAGGGACCGAAGUCGUGAAGCCGGGGG CUUCAGUCAAGCUGUCCUGCAAGGCCAGCGGCUAUAUCUUCACU AGCUACGACAUCGAUUGGGUGCGGCAGACUCCGGAGCAAGGACU CGAGUGGAUUGGGUGGAUCUUUCCGGGCGAGGGAUCAACCGAG UACAACGAAAAAUUUAAGGGACGGGCAACGCUGUCCGUGGACAA GAGCUCAUCUACGGCGUACAUGGAGCUGACGCGGCUCACGUCAG AGGAUUCCGCCGUCUACUUCUGUGCCAGGGGCGACUACUACCGG CGCUACUUUGAUCUGUGGGGACAAGGAACGACCGUGACUGUCUC AUCAGGCGGCGGCGGAUCGGGAGGAGGCGGAUCGGGUGGCGGU GGUUCGGACAUUCAGAUGACUCAAUCGCCCAGCUUCCUGUCGAC CUCACUGGGGAAUUCUAUUACGAUCACUUGUCACGCUUCGCAGA ACAUCAAGGGUUGGCUGGCUUGGUACCAGCAGAAAAGCGGUAAC GCCCCGCAACUGCUCAUCUACAAGGCAUCGUCCCUGCAAUCGGG AGUGCCGUCACGCUUUUCAGGAUCGGGCUCCGGAACCGAUUACA UCUUUACCAUCAGCAACCUGCAGCCGGAAGACAUCGCCACUUAC UACUGUCAACACUAUCAGAGCUUUCCGUGGACCUUUGGAGGGGG GACCAAAUUGGAGAUCAAGCGCGACUACAAGGAUGACGAUGACA AA Construct 4 AUGGAUUGGACCUGGAUCUUGUUUCUCGUCGCCGCAGCCACUCG 247 CGUUCAUAGCAAAGGAAUGUCAUACUCCAUGUGCACGGGAAAAU UCAAGGUGGUCAAAGAGAUCGCGGAGACUCAGCACGGCACCAUC GUCAUUCGCGUGCAAACUGAAGGAGAUGGAUCUCCCUGCAAGAU CCCGUUCGAGAUCAUGGACCUGGAAAAGAGACACGUCCUCGGUA GACUGAUCACCGUGAACCCGAUCGUGACGGAGAAGGAUUCCCCG GUGAAUAUUGAAGCAGAGCCUCCAUUUGGGGACUCAUACAUUA UCAUUGGGGUCGAGCCGGGCCAGCUGAAGCUGAAUUGGUUUAA GAAGGGCUCGUCAAUCGGACAGAUGUUCGAAACUACUAUGAGG GGUGCAAAGCGGAUGGCGAUCCUCUCGGGCGGAGAUAUCAUCAA ACUCCUUAACGAACAGGUGAACAAGGAGAUGCAGUCCUCAAACC UUUACAUGAGCAUGUCGUCCUGGUGUUACACCCAUAGCCUGGAC GGCGCUGGAUUGUUCCUGUUUGACCAUGCAGCGGAGGAAUACGA ACACGCCAAGAAGCUCAUCAUCUUCCUGAACGAGAAUAACGUGC CAGUGCAACUGACCUCCAUCUCGGCUCCUGAGCACAAGUUCGAA GGACUCACCCAGAUCUUCCAAAAGGCCUACGAACACGAACAGCA CAUCAGCGAAUCCAUCAACAAUAUCGUGGACCAUGCUAUCAAAA GCAAAGACCAUGCGACCUUCAACUUCCUGCAAUGGUAUGUCGCC GAACAGCACGAAGAGGAGGUGCUGUUCAAGGACAUUCUCGACAA AAUCGAAUUGAUAGGGAACGAAAAUCACGGUCUGUACCUGGCCG AUCAAUACGUGAAGGGAAUUGCCAAGUCGCGGAAGUCGU Dengue 2 prME AUGCUGAAUAUUCUGAACCGCCGCCGCCGGACUGCCGGGAUUAU 248 (Thailand/0168/1979) AAUUAUGAUGAUUCCCACCGUGAUGGCCUUCCACCUGACCACCC GGAACGGGGAACCACAUAUGAUCGUGUCCAGACAGGAGAAGGG AAAGUCCCUGCUGUUCAAGACCGAGGACGGCGUGAACAUGUGCA CCCUCAUGGCUAUGGACCUGGGCGAACUCUGCGAGGACACCAUC ACCUACAAGUGCCCCCUGUUGAGGCAGAACGAGCCGGAGGAUAU UGACUGCUGGUGCAAUUCGACCAGCACCUGGGUCACCUACGGGA CUUGCACCACAACCGGAGAACAUCGGCGCGAAAAGCGCAGCGUG GCUUUGGUGCCUCACGUCGGAAUGGGGCUGGAGACUAGAACCGA GACUUGGAUGUCGUCGGAAGGGGCCUGGAAACACGCACAGCGCA UCGAAACUUGGAUACUCAGGCAUCCCGGCUUCACCAUUAUGGCC GCGAUCCUGGCAUACACCAUCGGUACUACCCACUUCCAACGGGC CCUGAUCUUUAUCCUCCUGACCGCUGUCGCACCAUCCAUGACCA UGCGGUGUAUCGGUAUCAGCAACAGGGACUUCGUGGAGGGAGU GUCGGGAGGAUCCUGGGUGGAUAUUGUGCUGGAACACGGUUCC UGCGUCACUACCAUGGCGAAGAACAAGCCUACCCUGGACUUUGA GCUGAUCAAAACUGAGGCCAAGCAGCCGGCCACCCUGCGCAAGU ACUGCAUCGAAGCCAAGCUGACCAAUACCACUACCGAAUCCCGC UGUCCGACCCAAGGGGAGCCCUCCCUGAAUGAGGAGCAGGACAA GCGCUUCGUCUGCAAGCAUUCAAUGGUCGACCGCGGCUGGGGAA ACGGCUGCGGACUGUUCGGAAAGGGCGGCAUUGUGACCUGUGCC AUGUUCACUUGCAAGAAGAACAUGGAAGGAAAGAUCGUGCAGC CCGAAAACCUGGAGUAUACCAUCGUCGUGACCCCGCACUCCGGG GAAGAACACGCUGUGGGAAACGACACCGGAAAGCACGGAAAGGA GAUCAAAGUGACCCCACAGUCGAGCAUUACCGAGGCCGAACUUA CUGGUUACGGCACUGUGACGAUGGAAUGUUCACCGAGAACUGGA CUGGAUUUCAACGAAAUGGUGCUGCUCCAAAUGGAAAACAAGGC CUGGCUGGUGCACCGCCAGUGGUUUCUUGACCUCCCUCUCCCUU GGCUGCCUGGAGCAGACACUCAGGGUUCCAACUGGAUUCAGAAG GAAACACUCGUGACCUUCAAGAACCCUCACGCGAAGAAGCAGGA UGUGGUCGUGCUGGGAAGCCAGGAGGGAGCGAUGCAUACCGCCC UCACCGGCGCGACGGAGAUUCAGAUGUCCAGCGGAAACCUUCUG UUCACCGGACACCUGAAGUGCAGACUGAGGAUGGACAAGCUGCA GCUCAAGGGAAUGUCCUACUCCAUGUGCACUGGAAAGUUCAAGG UCGUGAAGGAGAUUGCCGAAACUCAGCAUGGUACCAUCGUGAUC CGGGUGCAAUAUGAAGGGGACGGAUCCCCGUGCAAGAUCCCUUU CGAAAUCAUGGACUUGGAGAAGCGACACGUGCUGGGCAGACUGA UCACAGUCAACCCCAUCGUGACUGAGAAGGAUUCACCCGUGAAC AUUGAAGCCGAGCCGCCUUUCGGCGAUAGCUACAUCAUCAUUGG CGUGGAACCGGGACAGCUUAAGCUCAACUGGUUCAAGAAGGGUU CCUCGAUCGGUCAAAUGUUUGAAACCACGAUGCGGGGUGCCAAA CGGAUGGCCAUUCUGGGAGACACCGCCUGGGAUUUCGGCUCCUU GGGCGGAGUGUUCACUUCUAUCGGAAAGGCGCUGCACCAAGUGU UCGGAGCCAUCUACGGCGCCGCGUUCUCGGGCGUCAGCUGGACC AUGAAGAUUCUGAUCGGGGUCAUCAUCACUUGGAUUGGGAUGA ACUCACGGUCCACCUCCCUGAGCGUGUCCCUUGUCCUGGUCGGC AUCGUGACCCUGUACCUCGGAGUGAUGGUGCAGGCUUAG Dengue 2 prME AUGCUUAACAUUCUCAACCGCCGCCGGAGAACUGCUGGUAUUAU 249 (Thailand/16681/1984) CAUUAUGAUGAUUCCCACUGUGAUGGCCUUCCACCUGACCACGC GGAACGGCGAACCCCAUAUGAUUGUCGGUCGGCAGGAAAAGGGG AAGUCCCUGCUGUUCAAAACUGAGGACGGAGUGAACAUGUGCAC CCUCAUGGCUAUUGACCUGGGAGAGCUGUGCGAAGAUACUAUCA CGUACAAGUGCCCCCUGCUGCGCCAGAACGAGCCUGAGGACAUU GACUGCUGGUGCAACUCCACGUCAACCUGGGUCACCUACGGAAC UUGCGCGACUACCGGCGAACAUCGCAGAGAAAAGAGAAGCGUGG CCCUCGUGCCGCACGUCGGGAUGGGGCUGGAAACCCGGACCGAA ACCUGGAUGUCCUCGGAAGGCGCCUGGAAGCACGUGCAGAGGAU CGAAACUUGGAUCCUCCGGCACCCGGGAUUCACCAUCAUGGCCG CCAUCCUCGCUUACACAAUCGGAACCACUCACUUUCAACGCGCC CUGAUCUUCAUCCUGCUUACCGCCGUGGCCCCGUCCAUGACCAU GCGCUGCAUUGGAAUGUCAAACCGGGACUUCGUCGAGGGAGUCU CCGGAGGAAGCUGGGUGGACAUCGUGCUGGAGCACGGCAGCUGU GUGACCACCAUGGCCAAGAACAAGCCAACUCUUGAUUUCGAACU GAUCAAGACCGAGGCCAAGCAGCCUGCCACUCUGAGGAAGUACU GUAUCGAAGCGAAGCUGACCAACACCACUACCGAAUCCCGCUGC CCGACCCAGGGCGAACCUUCCUUGAACGAAGAACAGGACAAGAG AUUCGUGUGCAAGCAUAGCAUGGUCGACAGGGGAUGGGGGAAC GGAUGUGGACUCUUUGGGAAGGGCGGAAUCGUCACCUGUGCGA UGUUCCGGUGCAAGAAGAACAUGGAGGGGAAGGUCGUGCAGCCC GAAAAUCUCGAGUACACUAUCGUGAUCACCCCGCAUUCCGGAGA GGAGCACGCCGUGGGCAACGACACCGGGAAGCACGGAAAGGAGA UCAAAAUUACCCCUCAAUCCUCCACCACCGAAGCCGAAUUGACU GGUUACGGUACCGUGACUAUGGAGUGCUCGCCGCGGACUGGCUU GGACUUCAACGAGAUGGUGCUGCUGCAAAUGGAGAACAAGGCCU GGCUGGUGCACCGGCAGUGGUUUCUUGAUCUGCCUCUGCCUUGG CUGCCCGGAGCCGACACCCAGGGUAGCAAUUGGAUCCAGAAAGA GACACUCGUGACCUUUAAGAACCCGCACGCAAAGAAGCAGGAUG UCGUGGUCCUGGGAAGCCAAGAAGGGGCAAUGCAUACCGCACUC ACUGGAGCCACUGAAAUCCAGAUGUCCUCCGGCAAUCUGCUGUU CACCGGCCAUCUGAAGUGCCGACUGCGCAUGGACAAGCUCCAGC UUAAGGGAAUGUCCUACUCCAUGUGUACUGGAAAGUUCAAAGU CGUGAAGGAAAUUGCCGAAACCCAGCACGGCACCAUAGUGAUCC GGGUGCAGUACGAGGGCGACGGCUCACCCUGCAAAAUCCCGUUC GAGAUUAUGGAUCUCGAAAAGCGCCACGUGCUGGGCAGACUGAU UACCGUGAACCCUAUCGUGACCGAGAAGGAUUCCCCAGUGAACA UCGAGGCCGAACCGCCCUUCGGAGACUCGUAUAUCAUCAUCGGC GUGGAGCCCGGCCAGCUGAAGCUGAACUGGUUCAAGAAGGGGUC GAGCAUCGGCCAGAUGUUCGAGACUACCAUGCGCGGCGCGAAGA GGAUGGCGAUCCUGGGGGAUACCGCUUGGGACUUCGGUUCCCUC GGCGGGGUGUUCACCUCGAUUGGGAAGGCCCUCCACCAAGUGUU CGGUGCAAUCUACGGAGCGGCGUUCAGCGGAGUGUCGUGGACCA UGAAGAUUCUGAUCGGCGUGAUCAUCACCUGGAUUGGCAUGAAC UCCCGGUCUACUAGCCUGUCGGUGACCCUGGUGCUGGUCGGAAU CGUGACCUUGUACCUGGGAGUGAUGGUGCAAGCUUAG Dengue 2 prME AUGCUGAACAUCCUGAACCGCAGAAGGAGAACCGCCGGUAUUAU 250 (Jamaica/1409/1983) UAUUAUGAUGAUCCCCACCGUGAUGGCAUUCCACCUGACUACCC GCAACGGAGAGCCGCAUAUGAUCGUGGGCCGCCAGGAAAAGGGA AAGUCCCUGCUGUUCAAGACUGAGGACGGCGUGAACAUGUGCAC UCUCAUGGCCAUCGACCUCGGCGAACUGUGCGAGGACACCAUUA CUUACAAGUGCCCGCUGCUGAGACAGAACGAGCCUGAGGACAUC GACUGUUGGUGUAACUCGACCUCCACCUGGGUCACCUACGGAAC GUGCGCCACAACCGGAGAACACCGCCGGGAAAAGCGGAGCGUGG CUCUGGUGCCGCACGUCGGAAUGGGUCUGGAGACUAGAACCGAA ACCUGGAUGUCAUCCGAGGGGGCAUGGAAACAUGUGCAGCGAAU CGAGACUUGGAUCCUGAGACACCCGGGCUUCACUAUCAUGGCGG CCAUCCUUGCCUACACCAUUGGCACUACUCACUUCCAACGGGCG CUGAUCUUCAUACUGCUCACCGCGGUGGCCCCCUCCAUGACGAU GCGCUGCAUCGGAAUCUCCAACCGGGACUUCGUGGAGGGCGUCA GCGGAGGCAGCUGGGUGGACAUCGUGUUGGAGCACGGAAGCUGC GUGACCACCAUGGCCAAGAACAAGCCCACUCUUGAUUUUGAGCU GAUCAAGACGGAAGCAAAGCAGCCGGCCACUCUGAGGAAGUACU GCAUCGAGGCCAAGCUCACCAACACAACCACCGAAUCUCGGUGC CCGACCCAAGGAGAGCCAUCACUGAACGAGGAACAGGACAAGAG AUUCCUGUGCAAACAUUCGAUGGUGGACAGGGGAUGGGGAAAU GGUUGCGGCCUGUUCGGCAAAGGAGGCAUUGUGACCUGUGCGAU GUUCACUUGCAAGAAAAACAUGGAGGGGAAGGUCGUGUUGCCG GAGAACCUGGAGUACACUAUCGUGAUUACCCCGCACUCCGGGGA GGAACAUGCCGUGGGAAAUGACACCGGAAAGCACGGGAAGGAA AUCAAAAUCACGCCUCAGUCCUCAAUCACCGAAGCCGAGCUUAC CGGCUACGGUACCGUGACCAUGGAGUGCAGCCCUCGGACUGGAC UGGACUUCAACGAGAUGGUGCUGCUGCAAAUGGAAGAUAAGGC CUGGCUGGUGCACCGGCAGUGGUUCUUGGAUUUGCCACUGCCUU GGCUGCCCGGCGCGGAUACCCAGGGUUCCAACUGGAUUCAGAAG GAAACCCUCGUGACCUUCAAGAAUCCUCACGCCAAGAAGCAGGA CGUGGUGGUGCUGGGUUCCCAAGAAGGGGCCAUGCAUACUGCCC UCACUGGAGCGACCGAAAUCCAGAUGUCGUCCGGCAACCUCCUG UUCACCGGCCACCUGAAGUGCCGCCUGCGGAUGGACAAGUUGCA GCUGAAGGGAAUGAGCUACUCGAUGUGUACCGGAAAGUUCAAG AUCGUGAAGGAAAUCGCCGAAACCCAGCACGGAACCAUCGUCAU UAGAGUGCAGUACGAAGGGGACGGCAGCCCGUGCAAGAUCCCCU UCGAAAUUAUGGACCUGGAGAAGCGCCACGUGCUCGGAAGGCUC AUCACUGUCAACCCAAUCGUCACCGAAAAGGACUCCCCUGUGAA CAUCGAAGCAGAGCCCCCUUUCGGGGACUCCUACAUUAUUAUCG GCGUGGAGCCCGGCCAGCUGAAGCUGAACUGGUUCAAGAAGGGA UCCUCGAUCGGACAGAUGUUCGAAACCACCAUGCGGGGAGCCAA GCGGAUGGCUAUUCUGGGAGAUACCGCUUGGGAUUUCGGCUCCC UCGGCGGCGUCUUUACUUCCAUCGGGAAAGCGCUCCACCAAGUG UUUGGAGCCAUCUACGGUGCCGCUUUUUCCGGGGUGUCAUGGAC CAUGAAGAUUCUUAUCGGGGUCAUUAUUACUUGGAUCGGCAUG AACUCCCGGAGCACCUCGCUGUCCGUGAGCCUCGUGCUCGUGGG GGUGGUCACUCUGUAUCUUGGUGCCAUGGUGCAGGCCUAG Dengue 2 prME AUGCUUAACAUCCUGAAUAGAAGAAGAAGAACCGCCGGCAUUAU 251 (Thailand/NGS- CAUUAUGAUGAUACCCACCGUGAUGGCCUUCCACCUGACUACUC C/1944) GCAACGGAGAGCCUCAUAUGAUCGUGUCGCGGCAGGAGAAGGGA AAGUCCCUGCUGUUUAAGACGGAGGACGGCGUGAACAUGUGCAC UCUUAUGGCAAUGGACCUUGGAGAGCUGUGCGAGGAUACCAUCA CCUACAAGUGUCCGUUCCUGAAGCAAAACGAGCCUGAGGAUAUU GACUGCUGGUGCAACUCCACCUCAACCUGGGUCACAUAUGGGAC CUGUACCACUACUGGCGAACACCGCCGCGAGAAAAGAAGCGUGG CGUUGGUGCCUCACGUCGGCAUGGGUCUGGAAACUCGGACCGAA ACUUGGAUGAGCUCAGAGGGGGCAUGGAAGCACGCCCAGAGGAU UGAAACCUGGAUUCUGCGCCACCCUGGAUUCACCAUCAUGGCGG CUAUUCUGGCGUACACUAUUGGAACCACCCACUUUCAGCGGGCC CUUAUCUUCAUCCUCCUCACUGCCGUGGCGCCCUCCAUGACUAU GCGGUGUAUCGGAAUUUCCAACCGCGACUUCGUGGAAGGAGUGU CCGGAGGCUCCUGGGUCGACAUUGUGCUGGAACAUGGUUCAUGC GUGACCACGAUGGCCAAGAACAAGCCCACCCUCGACUUCGAGCU GAUCGAGACUGAAGCCAAGCAGCCGGCCACUCUGCGGAAGUACU GUAUCGAGGCCAAGCUCACCAACACCACCACCGAUUCCCGCUGC CCGACCCAAGGAGAACCUUCGCUCAACGAGGAGCAGGACAAGCG GUUCGUGUGCAAGCACAGCAUGGUCGACAGGGGAUGGGGGAAU GGAUGCGGUCUGUUCGGAAAGGGAGGCAUUGUGACUUGUGCAA UGUUCACUUGCAAGAAGAACAUGAAGGGGAAGGUCGUGCAGCC GGAAAACCUGGAGUACACCAUCGUGAUCACCCCUCAUUCGGGCG AAGAACACGCUGUGGGGAAUGAUACCGGAAAGCACGGAAAGGA AAUUAAGAUCACACCCCAAUCCAGCAUCACUGAGGCAGAACUGA CCGGCUACGGCACUGUGACCAUGGAGUGCUCGCCUCGGACUGGC CUGGACUUCAACGAGAUGGUGCUGCUCCAAAUGGAAAACAAGGC CUGGCUGGUGCACAGACAGUGGUUCCUCGAUUUGCCCUUGCCGU GGCUCCCUGGCGCCGACACCCAGGGAUCUAACUGGAUCCAGAAG GAAACCCUUGUGACCUUCAAGAACCCGCACGCUAAGAAACAGGA UGUGGUGGUGCUGGGAAGCCAGGAAGGAGCAAUGCAUACCGCGC UCACGGGUGCCACCGAGAUCCAGAUGAGCUCCGGGAACCUCCUG UUCACCGGUCACCUGAAGUGCCGACUCCGCAUGGACAAACUGCA GCUCAAGGGGAUGUCCUACUCCAUGUGCACCGGGAAAUUCAAGG UCGUGAAGGAGAUCGCUGAGACUCAGCACGGUACUAUCGUGAUC CGGGUGCAGUAUGAGGGAGAUGGGAGCCCGUGCAAAAUCCCAUU UGAGAUCAUGGACUUGGAAAAGCGCCAUGUGCUGGGUCGGCUG AUUACCGUGAACCCAAUCGUCACCGAAAAGGACAGCCCCGUCAA CAUUGAAGCCGAACCACCCUUCGGAGACUCGUACAUCAUCAUUG GCGUGGAACCGGGCCAGCUGAAGCUGAACUGGUUCAAAAAGGGG UCCUCUAUCGGCCAAAUGAUCGAAACCACCAUGCGGGGAGCUAA GCGGAUGGCGAUUUUGGGAGACACUGCGUGGGACUUUGGCUCAC UGGGGGGAGUGUUCACCAGCAUCGGCAAAGCCCUGCACCAAGUG UUCGGUGCCAUCUACGGAGCCGCCUUCAGCGGAGUGUCCUGGAU CAUGAAGAUCCUGAUCGGCGUGAUCAUUACCUGGAUCGGCAUGA ACUCCAGGUCCACCUCGCUCUCCGUGUCGCUGGUGCUGGUCGGG GUCGUGACCCUGUACCUGGGAGUGAUGGUCCAGGCCUGA Dengue 2 prME AUGUUGAAUAUCCUGAACCGCCGCCGGAGAACUGCCGGAAUUAU 252 (PuertoRico/PR159- CAUUAUGAUGAUCCCUACCGUGAUGGCGUUCCACCUUACUACCC S1/1969) GGAACGGGGAGCCUCACAUGAUCGUGUCACGCCAGGAGAAGGGG AAAUCCCUGCUGUUCAAGACCAAGGACGGUACCAACAUGUGUAC CCUGAUGGCGAUGGACCUCGGAGAGCUGUGCGAGGACACCAUCA CCUACAAAUGCCCGUUCCUGAAGCAGAACGAGCCGGAAGAUAUU GACUGUUGGUGCAACUCCACCUCCACUUGGGUCACCUACGGAAC UUGCACCACUACUGGGGAGCAUAGACGGGAGAAGCGCUCCGUGG CCCUGGUGCCGCACGUCGGCAUGGGACUGGAAACCAGAACCGAG ACUUGGAUGUCCAGCGAAGGCGCCUGGAAGCACGCCCAGCGGAU UGAAACUUGGAUCCUGAGGCACCCGGGUUUUACCAUUAUGGCCG CUAUCUUGGCAUACACCAUCGGCACCACCCACUUCCAACGCGUC CUGAUCUUCAUCCUGCUGACCGCCAUUGCGCCCUCCAUGACCAU GCGGUGCAUCGGAAUCAGCAACCGCGACUUCGUGGAAGGCGUCA GCGGCGGUUCUUGGGUGGACAUCGUGUUGGAGCACGGAUCGUGC GUGACCACCAUGGCCAAGAACAAGCCGACCCUCGAUUUCGAGCU GAUCAAGACUGAAGCCAAGCAGCCAGCUACCCUGCGGAAGUAUU GCAUCGAAGCCAAGCUCACUAAUACUACGACCGACAGCCGGUGU CCGACCCAAGGAGAGCCCACCCUGAAUGAGGAACAAGACAAGCG CUUCGUGUGCAAGCAUUCCAUGGUGGACCGGGGCUGGGGAAACG GCUGCGGACUGUUCGGGAAAGGAGGAAUUGUGACUUGCGCCAU GUUCACUUGCAAGAAGAACAUGGAGGGGAAGAUCGUCCAGCCUG AGAACCUCGAGUACACGGUCGUGAUUACUCCGCACUCGGGAGAA GAACACGCCGUGGGCAACGACACCGGAAAGCAUGGGAAGGAAGU GAAAAUCACGCCCCAAUCGUCGAUUACCGAGGCUGAGCUGACCG GCUACGGCACCGUGACCAUGGAGUGCUCCCCGAGGACCGGACUG GACUUCAACGAAAUGGUGCUGCUGCAGAUGAAGGACAAGGCCUG GCUGGUGCACCGCCAGUGGUUCCUCGACCUCCCACUCCCCUGGC UGCCCGGAGCGGAUACGCAGGGAUCCAACUGGAUCCAGAAGGAA ACUCUUGUGACCUUCAAGAACCCUCAUGCCAAGAAGCAGGACGU GGUGGUCCUGGGAUCCCAAGAGGGCGCGAUGCACACCGCACUGA CCGGCGCCACCGAAAUUCAGAUGUCCUCCGGAAACCUCCUGUUC ACUGGCCACCUGAAGUGCAGACUCCGCAUGGACAAGCUGCAGCU CAAGGGGAUGAGCUACUCCAUGUGUACCGGAAAAUUCAAGGUCG UGAAGGAAAUUGCAGAAACACAGCAUGGGACAAUUGUCAUUCG GGUCCAGUACGAGGGCGAUGGUUCACCGUGCAAGACUCCAUUCG AGAUCAUGGAUCUGGAGAAAAGACACGUGCUGGGUCGGCUGAC UACCGUGAACCCAAUCGUGACUGAGAAGGACUCCCCCGUGAACA UCGAAGCCGAGCCUCCUUUUGGCGAUUCCUACAUCAUCAUUGGA GUGGAACCCGGACAGCUUAAGUUGGAUUGGUUCAAGAAGGGCU CCUCGAUCGGACAGAUGUUCGAAACCACCAUGCGCGGUGCCAAG CGAAUGGCCAUCCUGGGGGACACCGCCUGGGACUUCGGUAGCCU GGGCGGAGUGUUUACCUCAAUUGGAAAGGCUCUGCACCAAGUGU UUGGGGCGAUCUACGGAGCGGCCUUCAGCGGUGUCUCCUGGACU AUGAAGAUUCUCAUCGGAGUGAUAAUCACCUGGAUCGGCAUGA ACAGCCGGUCAACCAGCCUGUCCGUGUCCCUGGUGCUGGUCGGC AUCGUGACUCUCUACCUCGGAGUGAUGGUGCAGGCCUAG Dengue 2 prME AUGCUCAACAUACUGAACAGACGGAGAAGGACCGCCGGUAUUAU 253 (16681-PDK53) UAUCAUGAUGAUCCCUACUGUGAUGGCAUUCCACCUGACAACCC GCAACGGAGAGCCCCACAUGAUCGUGUCACGCCAGGAGAAAGGG AAGUCACUGCUGUUCAAGACCGAAGUCGGCGUGAACAUGUGUAC CCUGAUGGCGAUGGAUCUUGGCGAACUGUGCGAGGACACCAUCA CGUACAAGUGCCCCCUGUUGCGGCAAAACGAACCAGAGGACAUC GACUGCUGGUGUAACUCCACCUCGACCUGGGUCACCUACGGAAC CUGUACCACUAUGGGGGAACACCGGCGGGAGAAGCGCUCCGUGG CGCUCGUGCCUCAUGUCGGCAUGGGACUGGAGACUCGGACUGAA ACCUGGAUGUCGUCGGAGGGGGCCUGGAAGCACGUCCAGCGGAU CGAGACUUGGAUCCUUCGCCAUCCGGGCUUCACCAUGAUGGCCG CCAUCCUGGCCUACACCAUCGGAACCACCCAUUUCCAACGGGCC CUGAUCCUGAUCCUGUUGACUGCCGUGACCCCCUCCAUGACUAU GCGGUGCAUUGGGAUGUCGAACAGGGAUUUCGUGGAGGGAGUC AGCGGUGGCAGCUGGGUGGACAUCGUGCUGGAACAUGGAUCCUG CGUGACUACCAUGGCAAAGAACAAGCCAACCCUCGAUUUCGAAC UGAUCAAGACCGAGGCGAAACAGCCGGCGACCCUGAGGAAGUAC UGCAUCGAGGCCAAGCUCACCAACACCACUACCGAGAGCAGAUG CCCUACCCAAGGGGAACCUUCCCUGAACGAGGAGCAGGACAAGA GAUUCGUCUGCAAGCACUCCAUGGUGGACCGCGGCUGGGGAAAC GGAUGCGGACUCUUCGGAAAGGGCGGUAUUGUGACCUGUGCCAU GUUCCGCUGCAAGAAAAACAUGGAAGGGAAAGUGGUGCAGCCCG AGAACCUCGAGUACACUAUCGUGAUCACACCGCACAGCGGAGAA GAACACGCCGUGGGCAACGACACUGGAAAGCACGGGAAGGAAAU CAAGAUCACCCCGCAAUCCUCAAUCACUGAGGCUGAGUUGACCG GCUACGGGACUAUUACCAUGGAAUGCUCCCCACGAACGGGACUG GACUUCAACGAAAUUGUGUUGCUCCAAAUGGAAAACAAGGCCUG GCUCGUGCACCGGCAGUGGUUCCUGGAUCUGCCCCUGCCGUGGC UGCCGGGUGCCGACACUCAGGGGAGCAACUGGAUUCAGAAGGAA ACCCUUGUGACCUUCAAGAACCCCCACGCAAAGAAGCAGGACGU GGUGGUGCUGGGUAGCCAAGAAGGCGCCAUGCACACGGCCCUGA CCGGAGCGACCGAGAUCCAGAUGUCCAGCGGAAAUCUGCUCUUU ACUGGUCAUCUGAAGUGCAGACUUCGGAUGGACAAGCUGCAACU GAAGGGAAUGUCCUACUCAAUGUGCACUGGAAAGUUCAAGGUC GUGAAGGAGAUCGCCGAAACCCAGCACGGGACUAUCGUCAUCCG CGUGCAGUACGAAGGAGAUGGCUCCCCGUGCAAGAUCCCUUUCG AAAUCAUGGACCUGGAGAAGCGCCACGUGUUGGGGCGCCUUAUU ACUGUGAACCCCAUCGUGACCGAGAAGGACUCCCCUGUCAACAU CGAGGCUGAACCGCCAUUCGGAGAUUCCUAUAUCAUUAUCGGAG UGGAACCGGGCCAGCUCAAGCUGAAUUGGUUCAAGAAGGGAUCC UCGAUUGGCCAGAUGUUCGAAACGACUAUGCGGGGCGCUAAGCG CAUGGCCAUCCUGGGCGAUACUGCCUGGGAUUUUGGUUCUCUGG GCGGAGUGUUCACCUCCAUUGGAAAGGCCCUGCACCAAGUGUUC GGCGCCAUCUACGGUGCCGCGUUUAGCGGUGUCUCAUGGACCAU GAAAAUCCUCAUUGGCGUGAUCAUUACCUGGAUUGGCAUGAACU CCAGAAGCACUUCCCUGUCCGUGACCCUGGUGCUCGUCGGAAUU GUGACACUCUACCUCGGAGUGAUGGUGCAGGCUUGA Dengue 2 prME AUGCUGAACAUUUUGAACAGACGCCGAAGGACCGCAGGCAUUAU 254 (Peru/IQT2913/1996) CAUUAUGAUGAUCCCUACCGUGAUGGCCUUCCAUCUGACUACUA GGAACGGAGAGCCACAUAUGAUCGUGUCGCGCCAGGAAAAGGGA AAGAGCCUGCUUUUUAAAACCAAGGACGGCACGAACAUGUGCAC CCUUAUGGCCAUGGACCUGGGGGAGUUGUGCGAGGACACCAUCA CCUACAAGUGCCCGUUCCUGAAGCAAAACGAGCCCGAAGAUAUU GACUGCUGGUGCAACUCCACCUCCACCUGGGUCACUUAUGGGAC UUGCACCACCACCGGCGAACAUCGCAGAGAAAAGAGAAGCGUGG CCCUGGUCCCCCACGUCGGGAUGGGCCUCGAGACUCGGACCGAA ACUUGGAUGUCAUCAGAGGGCGCAUGGAAGCAUGCUCAGCGGAU CGAAACCUGGAUCCUGAGACACCCUGGUUUCACAAUUAUGGCCG CCAUUCUUGCGUACACGAUCGGAACGACUCAUUUCCAACGCGUG CUGAUCUUCAUUCUCCUGACCGCUAUUGCGCCGUCCAUGACUAU GCGGUGCAUCGGAAUCUCAAACCGGGACUUCGUGGAAGGAGUGU CGGGAGGAUCCUGGGUGGACAUUGUGCUGGAGCACGGUUCCUGC GUCACCACCAUGGCCAAAAACAAGCCUACCCUGGACUUCGAGCU GAUCAAGACUGAGGCCAAGCAGCCCGCGACCCUCCGGAAGUACU GCAUCGAGGCCAAGUUGACCAACACUACUACCGAUUCCCGGUGC CCGACCCAAGGAGAACCAACUCUGAACGAAGAACAGGAUAAGCG GUUUGUGUGCAAGCACUCAAUGGUGGACAGGGGAUGGGGCAAC GGCUGUGGACUGUUCGGAAAGGGUGGUAUUGUGACCUGUGCAA UGUUUACCUGUAAAAAGAAUAUGGAGGGGAAGAUCGUGCAGCC UGAAAAUCUCGAGUACACUGUCGUCAUCACCCCGCACUCGGGAG AGGAGCACGCUGUGGGCAACGACACCGGAAAGCACGGAAAGGAG GUCAAGAUAACCCCGCAAUCCUCCAUUACGGAAGCCGAACUGAC UGGUUACGGCACCGUGACUAUGGAGUGCUCCCCUCGGACCGGCC UGGACUUCAACGAAAUGGUGCUGCUCCAAAUGGAAGAUAAGGCC UGGCUGGUGCACAGGCAGUGGUUCCUGGAUCUCCCGCUGCCGUG GCUGCCUGGCGCUGACACUCAGGGAAGCAACUGGAUCCAGAAGG AAACCCUCGUGACCUUUAAGAACCCCCACGCCAAGAAGCAGGAU GUGGUGGUGUUGGGAAGCCAGGAGGGGGCCAUGCAUACUGCCCU CACCGGCGCGACCGAAAUCCAGAUGUCGUCCGGCAAUCUGCUGU UCACCGGACACCUCAAGUGUCGCCUUCGGAUGGACAAGCUGCAG CUGAAGGGAAUGAGCUACAGCAUGUGCACCGGGAAGUUCAAGA UCGUGAAGGAAAUCGCCGAAACCCAGCACGGAACCAUCGUGAUC CGGGUGCAGUACGAGGGCGACGGUUCUCCCUGCAAAAUCCCCUU CGAAAUCAUGGAUCUGGAGAAGAGACACGUCCUGGGUCGCCUGA UCACCGUGAACCCCAUUGUGACUGAGAAGGACUCCCCAGUGAAC AUCGAAGCGGAGCCCCCAUUCGGAGACAGCUACAUUAUCAUUGG UGCCGAACCGGGGCAGCUGAAACUGGACUGGUUCAAGAAGGGCA GCUCGAUUGGCCAAAUGUUCGAAACGACAAUGCGGGGCGCAAAG CGCAUGGCCAUCCUGGGAGACACUGCCUGGGACUUCGGGUCCCU UGGGGGGGUGUUCACCUCGAUCGGAAAAGCCUUGCACCAAGUGU UCGGCGCAAUCUACGGCGCCGCGUUCUCGGGAGUCUCCUGGACU AUGAAGAUCCUGAUCGGUGUCAUCAUCACCUGGAUCGGGAUGAA CUCCCGGUCCACUUCCCUCUCGGUGUCACUCGUGCUUGUGGGAA UUGUCACCCUGUACCUCGGAGUGAUGGUGCAGGCCUGA Dengue 2 prME AUGCUGAAUAUUCUGAACCGACGCCGCCGCACUGCCGGAAUCAU 255 (Thailand/PUO- UAUCAUGAUGAUCCCUACCGUGAUGGCGUUCCAUCUCACCACUC 218/1980) GGAAUGGCGAACCCCAUAUGAUCGUGUCGAGACAGGAAAAGGG AAAGAGCCUUUUGUUCAAAACUGAAGAUGGAGUGAACAUGUGC ACUCUCAUGGCAAUGGAUCUGGGCGAACUGUGCGAAGAUACCAU CACUUACAAGUGUCCGCUGUUGAGACAGAACGAGCCUGAGGACA UCGACUGCUGGUGUAACAGCACUUCCACCUGGGUCACCUACGGC ACUUGCACUACCACCGGAGAACACCGGCGCGAGAAGAGGAGCGU GGCUCUUGUGCCGCACGUCGGCAUGGGACUCGAGACUCGGACCG AAACCUGGAUGUCAUCCGAAGGAGCCUGGAAACACGCCCAACGG AUCGAAAUUUGGAUCCUGAGACACCCCGGUUUCACUAUCAUGGC CGCAAUCCUGGCGUACACUAUUGGCACCACGCACUUCCAGAGGG CCCUCAUUUUCAUCCUCCUGACUGCCGUGGCGCCAUCCAUGACC AUGAGAUGUAUUGGCAUUUCCAACCGCGAUUUCGUGGAGGGAG UGUCCGGAGGAUCCUGGGUCGACAUCGUGCUGGAACACGGAUCU UGCGUCACCACCAUGGCUAAGAACAAGCCCACCCUCGACUUCGA GCUGAUCAAGACAGAAGCCAAGCAGCCGGCCACCCUCCGCAAGU AUUGCAUUGAAGCCAAGCUUACCAACACCACCACCGAGUCGCGG UGCCCAACCCAAGGAGAGCCGAGCCUCAAUGAGGAACAGGACAA GCGCUUCGUGUGCAAACACAGCAUGGUCGACCGGGGUUGGGGCA ACGGAUGUGGCCUGUUCGGGAAGGGUGGCAUUGUGACUUGCGC AAUGUUCACUUGCAAGAAGAACAUGGAGGGGAAAGUGGUGCAA CCCGAGAACCUGGAGUACACCAUCGUCGUGACCCCACACUCCGG AGAGGAGCACGCCGUGGGAAACGACACGGGGAAGCAUGGAAAG GAGAUCAAGGUCACACCCCAAUCAUCUAUUACCGAGGCCGAACU GACCGGAUACGGUACUGUGACGAUGGAGUGCAGCCCGAGGACUG GACUGGACUUCAACGAAAUGGUGCUGCUGCAAAUGGAGAACAA GGCCUGGCUCGUGCACCGGCAGUGGUUUCUGGAUCUCCCACUGC CGUGGUUGCCGGGAGCCGACACCCAGGGGUCGAACUGGAUCCAG AAGGAAACUCUUGUGACGUUUAAGAAUCCUCACGCGAAGAAGCA GGACGUGGUGGUCCUGGGAUCGCAGGAAGGAGCUAUGCACACCG CUCUGACCGGCGCCACUGAGAUCCAGAUGUCCUCGGGCAACCUC CUGUUCACCGGUCAUCUGAAGUGCCGGCUGCGGAUGGACAAAUU GCAGCUGAAGGGGAUGUCCUACUCCAUGUGCACCGGGAAGUUCA AGGUCGUGAAGGAGAUCGCGGAAACUCAGCACGGCACCAUUGUC AUUAGAGUGCAGUACGAGGGAGAUGGUUCACCGUGCAAGAUAC CGUUCGAAAUCAUGGACCUGGAAAAGAGACAUGUCUUGGGACGC CUGAUCACUGUGAACCCUAUCGUGACCGAAAAGGACUCCCCUGU GAACAUCGAGGCGGAGCCGCCUUUCGGCGACUCCUACAUCAUUA UCGGAGUGGAGCCCGGGCAGCUGAAGCUCAACUGGUUUAAGAAG GGGUCCAGCAUCGGCCAGAUGUUCGAAACCACCAUGCGGGGGGC GAAGAGGAUGGCGAUCCUGGGAGACACCGCCUGGGAUUUCGGUU CACUGGGCGGAGUGUUCACCUCCAUCGGAAAGGCCCUGCACCAA GUGUUCGGCGCAAUCUACGGUGCUGCCUUCUCGGGAGUCUCCUG GACCAUGAAGAUCCUGAUCGGCGUGAUUAUCACAUGGAUCGGCA UGAACAGCCGGUCAACCUCCCUUUCCGUGUCCCUGGUGCUGGUC GGCAUCGUGACUCUGUACCUGGGCGUGAUGGUGCAGGCCUGA Dengue 2 prME AUGCUGAACAUUCUGAACCGGAGAAGAAGAACCGCCGGCAUUAU 256 (D2Y98P) with UAUCAUGAUGAUUCCCACUGUGAUGGCAUUUCACCUGACCACCC native leader GGAACGGAGAACCUCAUAUGAUCGUGUCGAGACAGGAGAAGGG AAAGUCCCUGCUGUUCAAGACAGAAAACGGAGUGAACAUGUGCA CCCUGAUGGCCAUGGAUCUCGGCGAACUGUGCGAGGAUACUAUC ACCUACAACUGUCCGUUGCUGCGCCAAAACGAGCCGGAGGACAU CGACUGCUGGUGUAACUCCACGUCGACCUGGGUCACCUACGGCA CUUGCACCGCGACCGGCGAACACAGAAGAGAGAAACGCUCCGUC GCUCUGGUGCCGCACGUCGGGAUGGGGCUUGAAACCCGGACUGA AACCUGGAUGAGCUCGGAGGGCGCUUGGAAGCAUGCCCAGCGCA UCGAAACUUGGGUGCUGAGGCAUCCAGGCUUCACAAUCAUGGCC GCCAUCCUCGCGUACACCAUCGGUACUACGUACUUCCAGCGGGU GUUGAUCUUCAUUCUGCUGACCGCCGUGGCCCCUAGCAUGACCA UGCGGUGCAUCGGGAUCUCCAACCGCGAUUUCGUGGAGGGGGUG UCCGGUGGAAGCUGGGUGGACAUUGUGCUGGAGCACGGCUCGUG CGUGACCACCAUGGCCAAGAACAAGCCCACCCUUGAUUUUGAGC UGAUCAAGACCGAAGCGAAACACCCCGCGACCCUCCGGAAGUAC UGCAUUGAAGCCAAGCUCACCAACACUACCACGGCCUCCCGGUG CCCUACCCAAGGAGAACCUUCCUUGAACGAAGAACAGGACAAGC GCUUCGUGUGCAAGCAUUCAAUGGUGGACCGGGGCUGGGGAAA UGGCUGUGGCCUCUUCGGAAAAGGCGGAAUUGUGACUUGCGCAA UGUUCACUUGCAAGAAGAACAUGGAGGGAAAGAUUGUGCAGCC CGAGAACCUCGAGUACACUAUUGUCAUCACUCCCCACUCCGGCG AAGAAAACGCUGUCGGCAACGACACCGGAAAGCAUGGAAAGGAG AUCAAGGUCACCCCGCAAUCCUCAAUUACUGAGGCAGAACUGAC CGGUUACGGAACUGUGACUAUGGAGUGUUCCCCUCGCACCGGCC UCGAUUUCAACGAGAUGGUGCUGCUGCAAAUGGAGAACAAGGCC UGGCUGGUGCACCGGCAGUGGUUCCUCGAUUUGCCCCUGCCGUG GCUGCCGGGAGCCGACACUCAGGGAUCCAACUGGAUCCAGAAAG AAACCCUCGUGACCUUCAAAAACCCCCACGCGAAGAAGCAGGAC GUGGUGGUGCUGGGUUCCCAAGAAGGGGCGAUGCAUACCGCCCU GACUGGUGCUACCGAAAUCCAGAUGUCAAGCGGAAAUCUCCUGU UUACCGGUCACCUGAAGUGCAGGCUCCGGAUGGACAAGUUGCAG CUGAAGGGGAUGUCGUACAGCAUGUGUACUGGGAAGUUCAAGG UCGUGAAGGAGAUUGCCGAAACCCAGCACGGAACCAUAGUCAUC AGGGUCCAGUACGAGGGCGACGGCAGCCCUUGCAAGAUCCCGUU CGAGAUCAUGGAUCUGGAGAAGCGACACGUGCUGGGCCGGCUUA UCACUGUGAAUCCAAUCGUGACCGAGAAAGACUCGCCCGUGAAC AUCGAAGCCGAGCCGCCGUUCGGCGACUCAUACAUCAUCAUCGG CGUGGAACCAGGACAGCUGAAGCUGUCAUGGUUCAAGAAGGGU UCCAGCAUUGGUCAGAUGUUCGAAACAACGAUGCGCGGAGCCAA GCGCAUGGCUAUCCUUGGGGACACCGCCUGGGACUUCGGGUCGC UGGGAGGAGUGUUUACCAGCAUCGGAAAGGCCCUGCACCAAGUG UUCGGUGCCAUCUACGGAGCCGCAUUUUCCGGAGUGUCGUGGAC UAUGAAGAUUCUGAUCGGCGUCGUGAUUACCUGGAUCGGGAUG AACUCCAGGUCUACUUCCCUCUCCGUGAGCCUGGUGCUGGUCGG CGUGGUCACCCUGUAUCUGGGCGUGAUGGUCCAGGCUUAG

TABLE 29 DENY Amino Acid Sequences SEQ ID Name Sequence NO DEN-1 MNNQRKKTGRPSFNMLKRARNRVSTVSQLAKRFSKGLLSGQGPMKL 259 (NC_001477.1) VMAFIAFLRFLAIPPTAGILARWGSFKKNGAIKVLRGFKKEISNMLNI MNRRKRSVTMLLMLLPTALAFHLTTRGGEPHMIVSKQERGKSLLFKT SAGVNMCTLIAMDLGELCEDTMTYKCPRITETEPDDVDCWCNATET WVTYGTCSQTGEHRRDKRSVALAPHVGLGLETRTETWMSSEGAWK QIQKVETWALRHPGFTVIALFLAHAIGTSITQKGIIFILLMLVTPSMAM RCVGIGNRDFVEGLSGATWVDVVLEHGSCVTTMAKDKPTLDIELLKT EVTNPAVLRKLCIEAKISNTTTDSRCPTQGEATLVEEQDTNFVCRRTF VDRGWGNGCGLFGKGSLITCAKFKCVTKLEGKIVQYENLKYSVIVTV HTGDQHQVGNETTEHGTTATITPQAPTSEIQLTDYGALTLDCSPRTGL DFNEMVLLTMKKKSWLVHKQWFLDLPLPWTSGASTSQETWNRQDL LVTFKTAHAKKQEVVVLGSQEGAMHTALTGATEIQTSGTTTIFAGHL KCRLKMDKLILKGMSYVMCTGSFKLEKEVAETQHGTVLVQVKYEGT DAPCKIPFSSQDEKGVTQNGRLITANPIVTDKEKPVNIEAEPPFGESYIV VGAGEKALKLSWFKKGSSIGKMFEATARGARRMAILGDTAWDFGSI GGVFTSVGKLIHQIFGTAYGVLFSGVSWTMKIGIGILLTWLGLNSRSTS LSMTCIAVGMVTLYLGVMVQADSGCVINWKGRELKCGSGIFVTNEV HTWTEQYKFQADSPKRLSAAIGKAWEEGVCGIRSATRLENIMWKQIS NELNHILLENDMKFTVVVGDVSGILAQGKKMIRPQPMEHKYSWKSW GKAKIIGADVQNTTFIIDGPNTPECPDNQRAWNIWEVEDYGFGIFTTNI WLKLRDSYTQVCDHRLMSAAIKDSKAVHADMGYWIESEKNETWKL ARASFIEVKTCIWPKSHTLWSNGVLESEMIIPKIYGGPISQHNYRPGYF TQTAGPWHLGKLELDFDLCEGTTVVVDEHCGNRGPSLRTTTVTGKTI HEWCCRSCTLPPLRFKGEDGCWYGMEIRPVKEKEENLVKSMVSAGS GEVDSFSLGLLCISIMIEEVMRSRWSRKMLMTGTLAVFLLLTMGQLT WNDLIRLCIMVGANASDKMGMGTTYLALMATFRMRPMFAVGLLFR RLTSREVLLLTVGLSLVASVELPNSLEELGDGLAMGIMMLKLLTDFQS HQLWATLLSLTFVKTTFSLHYAWKTMAMILSIVSLFPLCLSTTSQKTT WLPVLLGSLGCKPLTMFLITENKIWGRKSWPLNEGIMAVGIVSILLSSL LKNDVPLAGPLIAGGMLIACYVISGSSADLSLEKAAEVSWEEEAEHSG ASHNILVEVQDDGTMKIKDEERDDTLTILLKATLLAISGVYPMSIPATL FVWYFWQKKKQRSGVLWDTPSPPEVERAVLDDGIYRILQRGLLGRS QVGVGVFQEGVFHTMWHVTRGAVLMYQGKRLEPSWASVKKDLISY GGGWRFQGSWNAGEEVQVIAVEPGKNPKNVQTAPGTFKTPEGEVGA IALDFKPGTSGSPIVNREGKIVGLYGNGVVTTSGTYVSAIAQAKASQE GPLPEIEDEVFRKRNLTIMDLHPGSGKTRRYLPAIVREAIKRKLRTLVL APTRVVASEMAEALKGMPIRYQTTAVKSEHTGKEIVDLMCHATFTM RLLSPVRVPNYNMIIMDEAHFTDPASIAARGYISTRVGMGEAAAIFMT ATPPGSVEAFPQSIQDEERDIPERSWNSGYDWITDFPGKTVWFVPSIKS GNDIANCLRKNGKRVVQLSRKTFDTEYQKTKNNDWDYVVTTDISEM GANFRADRVIDPRRCLKPVILKDGPERVILAGPMPVTVASAAQRRGRI GRNQNKEGDQYIYMGQPLNNDEDHAHWTEAKMLLDNINTPEGIIPAL FEPEREKSAAIDGEYRLRGEARKTFVELMRRGDLPVWLSYKVASEGF QYSDRRWCFDGERNNQVLEENMDVEIWTKEGERKKLRPRWLDART YSDPLALREFKEFAAGRRSVSGDLILEIGKLPQHLTQRAQNALDNLV MLHNSEQGGKAYRHAMEELPDTIETLMLLALIAVLTGGVTLFFLSGR GLGKTSIGLLCVIASSALLWMASVEPHWIAASIILEFFLMVLLIPEPDR QRTPQDNQLAYVVIGLLFMILTVAANEMGLLETTKKDLGIGHAAAEN HHHAAMLDVDLHPASAWTLYAVATTIITPMMRHTIENTTANISLTAIA NQAAILMGLDKGWPISKMDIGVPLLALGCYSQVNPLTLTAAVLMLV AHYAIIGPGLQAKATREAQKRTAAGIMKNPTVDGIVAIDLDPVVYDA KFEKQLGQIMLLILCTSQILLMRTTWALCESITLATGPLTTLWEGSPGK FWNTTIAVSMANIFRGSYLAGAGLAFSLMKSLGGGRRGTGAQGETLG EKWKRQLNQLSKSEFNTYKRSGIIEVDRSEAKEGLKRGETTKHAVSR GTAKLRWFVERNLVKPEGKVIDLGCGRGGWSYYCAGLKKVTEVKG YTKGGPGHEEPIPMATYGWNLVKLYSGKDVFFTPPEKCDTLLCDIGE SSPNPTIEEGRTLRVLKMVEPWLRGNQFCIKILNPYMPSVVETLEQMQ RKHGGMLVRNPLSRNSTHEMYWVSCGTGNIVSAVNMTSRMLLNRFT MAHRKPTYERDVDLGAGTRHVAVEPEVANLDIIGQRIENIKNEHKST WHYDEDNPYKTWAYHGSYEVKPSGSASSMVNGVVRLLTKPWDVIP MVTQIAMTDTTPFGQQRVFKEKVDTRTPKAKRGTAQIMEVTARWLW GFLSRNKKPRICTREEFTRKVRSNAAIGAVFVDENQWNSAKEAVEDE RFWDLVHRERELHKQGKCATCVYNMMGKREKKLGEFGKAKGSRAI WYMWLGARFLEFEALGFMNEDHWFSRENSLSGVEGEGLHKLGYILR DISKIPGGNMYADDTAGWDTRITEDDLQNEAKITDIMEPEHALLATSI FKLTYQNKVVRVQRPAKNGTVMDVISRRDQRGSGQVGTYGLNTFTN MEAQLIRQMESEGIFSPSELETPNLAERVLDWLKKHGTERLKRMAISG DDCVVKPIDDRFATALTALNDMGKVRKDIPQWEPSKGWNDWQQVP FCSHHFHQLIMKDGREIVVPCRNQDELVGRARVSQGAGWSLRETACL GKSYAQMWQLMYFHRRDLRLAANAICSAVPVDWVPTSRTTWSIHA HHQWMTTEDMLSVWNRVWIEENPWMEDKTHVSSWEDVPYLGKRE DQWCGSLIGLTARATWATNIQVAINQVRRLIGNENYLDFMTSMKRFK NESDPEGALW DEN-2 MNNQRKKAKNTPFNMLKRERNRVSTVQQLTKRFSLGMLQGRGPLKL 260 (NC_001474.2) FMALVAFLRFLTIPPTAGILKRWGTIKKSKAINVLRGFRKEIGRMLNIL NRRRRSAGMIIMLIPTVMAFHLTTRNGEPHMIVSRQEKGKSLLFKTED GVNMCTLMAMDLGELCEDTITYKCPLLRQNEPEDIDCWCNSTSTWV TYGTCTTMGEHRREKRSVALVPHVGMGLETRTETWMSSEGAWKHV QRIETWILRHPGFTMMAAILAYTIGTTHFQRALIFILLTAVTPSMTMRC IGMSNRDFVEGVSGGSWVDIVLEHGSCVTTMAKNKPTLDFELIKTEA KQPATLRKYCIEAKLTNTTTESRCPTQGEPSLNEEQDKRFVCKHSMV DRGWGNGCGLFGKGGIVTCAMFRCKKNMEGKVVQPENLEYTIVITP HSGEEHAVGNDTGKHGKEIKITPQSSITEAELTGYGTVTMECSPRTGL DFNEMVLLQMENKAWLVHRQWFLDLPLPWLPGADTQGSNWIQKET LVTFKNPHAKKQDVVVLGSQEGAMHTALTGATEIQMSSGNLLFTGH LKCRLRMDKLQLKGMSYSMCTGKFKVVKEIAETQHGTIVIRVQYEG DGSPCKIPFEIMDLEKRHVLGRLITVNPIVTEKDSPVNIEAEPPFGDSYII IGVEPGQLKLNWFKKGSSIGQMFETTMRGAKRMAILGDTAWDFGSL GGVFTSIGKALHQVFGAIYGAAFSGVSWTMKILIGVIITWIGMNSRSTS LSVTLVLVGIVTLYLGVMVQADSGCVVSWKNKELKCGSGIFITDNVH TWTEQYKFQPESPSKLASAIQKAHEEGICGIRSVTRLENLMWKQITPE LNHILSENEVKLTIMTGDIKGIMQAGKRSLRPQPTELKYSWKTWGKA KWILSTESHNQTFLIDGPETAECPNTNRAWNSLEVEDYGFGVFTTNIW LKLKEKQDVFCDSKLMSAAIKDNRAVHADMGYWIESALNDTWKIEK ASFIEVKNCHWPKSHTLWSNGVLESEMIIPKNLAGPVSQHNYRPGYH TQITGPWHLGKLEMDFDFCDGTTVVVTEDCGNRGPSLRTTTASGKLI TEWCCRSCTLPPLRYRGEDGCWYGMEIRPLKEKEENLVNSLVTAGH GQVDNFSLGVLGMALFLEEMLRTRVGTKHAILLVAVSFVTLITGNMS FRDLGRVMVMVGATMTDDIGMGVTYLALLAAFKVRPTFAAGLLLR KLTSKELMMTTIGIVLLSQSTIPETILELTDALALGMMVLKMVRNMEK YQLAVTIMAILCVPILQNAWKVSCTILAVVSVSPLLLTSSQQKTDWIPL ALTIKGLNPTAIFLTTLSRTSKKRSWPLNEAIMAVGMVSILASSLLKND IPMTGPLVAGGLLTVCYVLTGRSADLELERAADVKWEDQAEISGSSPI LSITISEDGSMSIKNEEEEQTLTILIRTGLLVISGLFPVSIPITAAAWYLW EVKKQRAGVLWDVPSPPPMGKAELEDGAYRIKQKGILGYSQIGAGV YKEGTFHTMWHVTRGAVLMHKGKRIEPSWADVKKDLISYGGGWKL EGEWKEGEEVQVLALEPGKNPRAVQTKPGLFKTNAGTIGAVSLDFSP GTSGSPIIDKKGKVVGLYGNGVVTRSGAYVSAIAQTEKSIEDNPEIED DIFRKRRLTIMDLHPGAGKTKRYLPAIVREAIKRGLRTLILAPTRVVAA EMEEALRGLPIRYQTPAIRAEHTGREIVDLMCHATFTMRLLSPVRVPN YNLIIMDEAHFTDPASIAARGYISTRVEMGEAAGIFMTATPPGSRDPFP QSNAPIIDEEREIPERSWNSGHEWVTDFKGKTVWFVPSIKAGNDIAAC LRKNGKKVIQLSRKTFDSEYVKTRTNDWDFVVTTDISEMGANFKAER VIDPRRCMKPVILTDGEERVILAGPMPVTHSSAAQRRGRIGRNPKNEN DQYIYMGEPLENDEDCAHWKEAKMLLDNINTPEGIIPSMFEPEREKV DAIDGEYRLRGEARKTFVDLMRRGDLPVWLAYRVAAEGINYADRR WCFDGVKNNQILEENVEVEIWTKEGERKKLKPRWLDARIYSDPLALK EFKEFAAGRKSLTLNLITEMGRLPTFMTQKARDALDNLAVLHTAEAG GRAYNHALSELPETLETLLLLTLLATVTGGIFLFLMSGRGIGKMTLGM CCIITASILLWYAQIQPHWIAASIILEFFLIVLLIPEPEKQRTPQDNQLTY VVIAILTVVAATMANEMGFLEKTKKDLGLGSIATQQPESNILDIDLRP ASAWTLYAVATTFVTPMLRHSIENSSVNVSLTAIANQATVLMGLGKG WPLSKMDIGVPLLAIGCYSQVNPITLTAALFLLVAHYAIIGPGLQAKA TREAQKRAAAGIMKNPTVDGITVIDLDPIPYDPKFEKQLGQVMLLVL CVTQVLMMRTTWALCEALTLATGPISTLWEGNPGRFWNTTIAVSMA NIFRGSYLAGAGLLFSIMKNTTNTRRGTGNIGETLGEKWKSRLNALG KSEFQIYKKSGIQEVDRTLAKEGIKR GETDHHAVSRGSAKLRWFVERNMVTPEGKVVDLGCGRGGWSYYCG GLKNVREVKGLTKGGPGHEEPIPMSTYGWNLVRLQSGVDVFFIPPEK CDTLLCDIGESSPNPTVEAGRTLRVLNLVENWLNNNTQFCIKVLNPY MPSVIEKMEALQRKYGGALVRNPLSRNSTHEMYWVSNASGNIVSSV NMISRMLINRFTMRYKKATYEPDVDLGSGTRNIGIESEIPNLDIIGKRIE KIKQEHETSWHYDQDHPYKTWAYHGSYETKQTGSASSMVNGVVRL LTKPWDVVPMVTQMAMTDTTPFGQQRVFKEKVDTRTQEPKEGTKK LMKITAEWLWKELGKKKTPRMCTREEFTRKVRSNAALGAIFTDENK WKSAREAVEDSRFWELVDKERNLHLEGKCETCVYNMMGKREKKLG EFGKAKGSRAIWYMWLGARFLEFEALGFLNEDHWFSRENSLSGVEG EGLHKLGYILRDVSKKEGGAMYADDTAGWDTRITLEDLKNEEMVTN HMEGEHKKLAEAIFKLTYQNKVVRVQRPTPRGTVMDIISRRDQRGSG QVGTYGLNTFTNMEAQLIRQMEGEGVFKSIQHLTITEEIAVQNWLAR VGRERLSRMAISGDDCVVKPLDDRFASALTALNDMGKIRKDIQQWEP SRGWNDWTQVPFCSHHFHELIMKDGRVLVVPCRNQDELIGRARISQG AGWSLRETACLGKSYAQMWSLMYFHRRDLRLAANAICSAVPSHWV PTSRTTWSIHAKHEWMTTEDMLTVWNRVWIQENPWMEDKTPVESW EEIPYLGKREDQWCGSLIGLTSRATWAKNIQAAINQVRSLIGNEEYTD YMPSMKRFRREEEEAGVLW DEN-3 MNNQRKKTGKPSINMLKRVRNRVSTGSQLAKRFSKGLLNGQGPMKL 261 (NC_001475.2) VMAFIAFLRFLAIPPTAGVLARWGTFKKSGAIKVLKGFKKEISNMLSII NQRKKTSLCLMMILPAALAFHLTSRDGEPRMIVGKNERGKSLLFKTA SGINMCTLIAMDLGEMCDDTVTYKCPHITEVEPEDIDCWCNLTSTWV TYGTCNQAGEHRRDKRSVALAPHVGMGLDTRTQTWMSAEGAWRQ VEKVETWALRHPGFTILALFLAHYIGTSLTQKVVIFILLMLVTPSMTM RCVGVGNRDFVEGLSGATWVDVVLEHGGCVTTMAKNKPTLDIELQK TEATQLATLRKLCIEGKITNITTDSRCPTQGEAVLPEEQDQNYVCKHT YVDRGWGNGCGLFGKGSLVTCAKFQCLEPIEGKVVQYENLKYTVIIT VHTGDQHQVGNETQGVTAEITPQASTTEAILPEYGTLGLECSPRTGLD FNEMILLTMKNKAWMVHRQWFFDLPLPWASGATTETPTWNRKELL VTFKNAHAKKQEVVVLGSQEGAMHTALTGATEIQNSGGTSIFAGHLK CRLKMDKLELKGMSYAMCTNTFVLKKEVSETQHGTILIKVEYKGED APCKIPFSTEDGQGKAHNGRLITANPVVTKKEEPVNIEAEPPFGESNIVI GIGDNALKINWYKKGSSIGKMFEATERGARRMAILGDTAWDFGSVG GVLNSLGKMVHQIFGSAYTALFSGVSWVMKIGIGVLLTWIGLNSKNT SMSFSCIAIGIITLYLGAVVQADMGCVINWKGKELKCGSGIFVTNEVH TWTEQYKFQADSPKRLATAIAGAWENGVCGIRSTTRMENLLWKQIA NELNYILWENNIKLTVVVGDTLGVLEQGKRTLTPQPMELKYSWKTW GKAKIVTAETQNSSFIIDGPNTPECPSASRAWNVWEVEDYGFGVFTTN IWLKLREVYTQLCDHRLMSAAVKDERAVHADMGYWIESQKNGSWK LEKASLIEVKTCTWPKSHTLWTNGVLESDMIIPKSLAGPISQHNYRPG YHTQTAGPWHLGKLELDFNYCEGTTVVITESCGTRGPSLRTTTVSGK LIHEWCCRSCTLPPLRYMGEDGCWYGMEIRPISEKEENMVKSLVSAG SGKVDNFTMGVLCLAILFEEVLRGKFGKKHMIAGVFFTFVLLLSGQIT WRDMAHTLIMIGSNASDRMGMGVTYLALIATFKIQPFLALGFFLRKL TSRENLLLGVGLAMATTLQLPEDIEQMANGVALGLMALKLITQFETY QLWTALVSLTCSNTIFTLTVAWRTATLILAGVSLLPVCQSSSMRKTD WLPMTVAAMGVPPLPLFIFSLKDTLKRRSWPLNEGVMAVGLVSILAS SLLRNDVPMAGPLVAGGLLIACYVITGTSADLTVEKAPDVTWEEEAE QTGVSHNLMITVDDDGTMRIKDDETENILTVLLKTALLIVSGIFPYSIP ATLLVWHTWQKQTQRSGVLWDVPSPPETQKAELEEGVYRIKQQGIF GKTQVGVGVQKEGVFHTMWHVTRGAVLTHNGKRLEPNWASVKKD LISYGGGWRLSAQWQKGEEVQVIAVEPGKNPKNFQTTPGTFQTTTGE IGAIALDFKPGTSGSPIINREGKVVGLYGNGVVTKNGGYVSGIAQTNA EPDGPTPELEEEMFKKRNLTIMDLHPGSGKTRKYLPAIVREAIKRRLR TLILAPTRVVAAEMEEALKGLPIRYQTTATKSEHTGREIVDLMCHATF TMRLLSPVRVPNYNLIIMDEAHFTDPASIAARGYISTRVGMGEAAAIF MTATPPGTADAFPQSNAPIQDEERDIPERSWNSGNEWITDFAGKTVW FVPSIKAGNDIANCLRKNGKKVIQLSRKTFDTEYQKTKLNDWDFVVT TDISEMGANFKADRVIDPRRCLKPVILTDGPERVILAGPMPVTAASAA QRRGRVGRNPQKENDQYIFTGQPLNNDEDHAHWTEAKMLLDNINTP EGIIPALFEPEREKSAAIDGEYRLKGESRKTFVELMRRGDLPVWLAHK VASEGIKYTDRKWCFDGQRNNQILEENMDVEIWTKEGEKKKLRPRW LDARTYSDPLALKEFKDFAAGRKSIALDLVTEIGRVPSHLAHRTRNAL DNLVMLHTSEDGGRAYRHAVEELPETMETLLLLGLMILLTGGAMLFL ISGKGIGKTSIGLICVIASSGMLWMAEVPLQWIASAIVLEFFMMVLLIP EPEKQRTPQDNQLAYVVIGILTLAATIAANEMGLLETTKRDLGMSKEP GVVSPTSYLDVDLHPASAWTLYAVATTVITPMLRHTIENSTANVSLA AIANQAVVLMGLDKGWPISKMDLGVPLLALGCYSQVNPLTLTAAVL LLITHYAIIGPGLQAKATREAQKRTAAGIMKNPTVDGIMTIDLDSVIFD SKFEKQLGQVMLLVLCAVQLLLMRTSWALCEALTLATGPITTLWEGS PGKFWNTTIAVSMANIFRGSYLAGAGLAFSIMKSVGTGKRGTGSQGE TLGEKWKKKLNQLSRKEFDLYKKSGITEVDRTEAKEGLKRGETTHH AVSRGSAKLQWFVERNMVVPEGRVIDLGCGRGGWSYYCAGLKKVT EVRGYTKGGPGHEEPVPMSTYGWNIVKLMSGKDVFYLPPEKCDTLL CDIGESSPSPTVEESRTIRVLKMVEPWLKNNQFCIKVLNPYMPTVIEHL ERLQRKHGGMLVRNPLSRNSTHEMYWISNGTGNIVSSVNMVSRLLL NRFTMTHRRPTIEKDVDLGAGTRHVNAEPETPNMDVIGERIKRIKEEH NSTWHYDDENPYKTWAYHGSYEVKATGSASSMINGVVKLLTKPWD VVPMVTQMAMTDTTPFGQQRVFKEKVDTRTPRPMPGTRKAMEITAE WLWRTLGRNKRPRLCTREEFTKKVRTNAAMGAVFTEENQWDSAKA AVEDEEFWKLVDRERELHKLGKCGSCVYNMMGKREKKLGEFGKAK GSRAIWYMWLGARYLEFEALGFLNEDHWFSRENSYSGVEGEGLHKL GYILRDISKIPGGAMYADDTAGWDTRITEDDLHNEEKIIQQMDPEHRQ LANAIFKLTYQNKVVKVQRPTPTGTVMDIISRKDQRGSGQLGTYGLN TFTNMEAQLVRQMEGEGVLTKADLENPHLLEKKITQWLETKGVERL KRMAISGDDCVVKPIDDRFANALLALNDMGKVRKDIPQWQPSKGWH DWQQVPFCSHHFHELIMKDGRKLVVPCRPQDELIGRARISQGAGWSL RETACLGKAYAQMWSLMYFHRRDLRLASNAICSAVPVHWVPTSRTT WSIHAHHQWMTTEDMLTVWNRVWIEENPWMEDKTPVTTWENVPY LGKREDQWCGSLIGLTSRATWAQNIPTAIQQVRSLIGNEEFLDYMPSM KRFRKEEESEGAIW DEN-4 MNQRKKVVRPPFNMLKRERNRVSTPQGLVKRFSTGLFSGKGPLRMV 262 (NC_002640.1) LAFITFLRVLSIPPTAGILKRWGQLKKNKAIKILIGFRKEIGRMLNILNG RKRSTITLLCLIPTVMAFSLSTRDGEPLMIVAKHERGRPLLFKTTEGIN KCTLIAMDLGEMCEDTVTYKCPLLVNTEPEDIDCWCNLTSTWVMYG TCTQSGERRREKRSVALTPHSGMGLETRAETWMSSEGAWKHAQRVE SWILRNPGFALLAGFMAYMIGQTGIQRTVFFVLMMLVAPSYGMRCV GVGNRDFVEGVSGGAWVDLVLEHGGCVTTMAQGKPTLDFELTKTT AKEVALLRTYCIEASISNITTATRCPTQGEPYLKEEQDQQYICRRDVV DRGWGNGCGLFGKGGVVTCAKFSCSGKITGNLVQIENLEYTVVVTV HNGDTHAVGNDTSNHGVTAMITPRSPSVEVKLPDYGELTLDCEPRSG IDFNEMILMKMKKKTWLVHKQWFLDLPLPWTAGADTSEVHWNYKE RMVTFKVPHAKRQDVTVLGSQEGAMHSALAGATEVDSGDGNHMFA GHLKCKVRMEKLRIKGMSYTMCSGKFSIDKEMAETQHGTTVVKVKY EGAGAPCKVPIEIRDVNKEKVVGRIISSTPLAENTNSVTNIELEPPFGDS YIVIGVGNSALTLHWFRKGSSIGKMFESTYRGAKRMAILGETAWDFG SVGGLFTSLGKAVHQVFGSVYTTMFGGVSWMIRILIGFLVLWIGTNSR NTSMAMTCIAVGGITLFLGFTVQADMGCVASWSGKELKCGSGIFVVD NVHTWTEQYKFQPESPARLASAILNAHKDGVCGIRSTTRLENVMWK QITNELNYVLWEGGHDLTVVAGDVKGVLTKGKRALTPPVSDLKYSW KTWGKAKIFTPEARNSTFLIDGPDTSECPNERRAWNSLEVEDYGFGM FTTNIWMKFREGSSEVCDHRLMSAAIKDQKAVHADMGYWIESSKNQ TWQIEKASLIEVKTCLWPKTHTLWSNGVLESQMLIPKSYAGPFSQHN YRQGYATQTVGPWHLGKLEIDFGECPGTTVTIQEDCDHRGPSLRTTT ASGKLVTQWCCRSCTMPPLRFLGEDGCWYGMEIRPLSEKEENMVKS QVTAGQGTSETFSMGLLCLTLFVEECLRRRVTRKHMILVVVITLCAIIL GGLTWMDLLRALIMLGDTMSGRIGGQIHLAIMAVFKMSPGYVLGVF LRKLTSRETALMVIGMAMTTVLSIPHDLMELIDGISLGLILLKIVTQFD NTQVGTLALSLTFIRSTMPLVMAWRTIMAVLFVVTLIPLCRTSCLQKQ SHWVEITALILGAQALPVYLMTLMKGASRRSWPLNEGIMAVGLVSLL GSALLKNDVPLAGPMVAGGLLLAAYVMSGSSADLSLEKAANVQWD EMADITGSSPIVEVKQDEDGSFSIRDVEETNMITLLVKLALITVSGLYP LAIPVTMTLWYMWQVKTQRSGALWDVPSPAATKKAALSEGVYRIM QRGLFGKTQVGVGIHMEGVFHTMWHVTRGSVICHETGRLEPSWADV RNDMISYGGGWRLGDKWDKEEDVQVLAIEPGKNPKHVQTKPGLFKT LTGEIGAVTLDFKPGTSGSPIINRKGKVIGLYGNGVVTKSGDYVSAITQ AERIGEPDYEVDEDIFRKKRLTIMDLHPGAGKTKRILPSIVREALKRRL RTLILAPTRVVAAEMEEALRGLPIRYQTPAVKSEHTGREIVDLMCHAT FTTRLLSSTRVPNYNLIVMDEAHFTDPSSVAARGYISTRVEMGEAAAI FMTATPPGATDPFPQSNSPIEDIEREIPERSWNTGFDWITDYQGKTVWF VPSIKAGNDIANCLRKSGKKVIQLSRKTFDTEYPKTKLTDWDFVVTTD ISEMGANFRAGRVIDPRRCLKPVILPDGPERVILAGPIPVTPASAAQRR GRIGRNPAQEDDQYVFSGDPLKNDEDHAHWTEAKMLLDNIYTPEGII PTLFGPEREKTQAIDGEFRLRGEQRKTFVELMRRGDLPVWLSYKVAS AGISYEDREWCFTGERNNQILEENMEVEIWTREGEKKKLRPRWLDAR VYADPMALKDFKEFASGRKSITLDILTEIASLPTYLSSRAKLALDNIVM LHTTERGGRAYQHALNELPESLETLMLVALLGAMTAGIFLFFMQGKG IGKLSMGLITIAVASGLLWVAEIQPQWIAASIILEFFLMVLLIPEPEKQR TPQDNQLIYVILTILTIIGLIAANEMGLIEKTKTDFGFYQVKTETTILDV DLRPASAWTLYAVATTILTPMLRHTIENTSANLSLAAIANQAAVLMG LGKGWPLHRMDLGVPLLAMGCYSQVNPTTLTASLVMLLVHYAIIGP GLQAKATREAQKRTAAGIMKNPTVDGITVIDLEPISYDPKFEKQLGQV MLLVLCAGQLLLMRTTWAFCEVLTLATGPILTLWEGNPGRFWNTTIA VSTANIFRGSYLAGAGLAFSLIKNAQTPRRGTGTTGETLGEKWKRQL NSLDRKEFEEYKRSGILEVDRTEAKSALKDGSKIKHAVSRGSSKIRWI VERGMVKPKGKVVDLGCGRGGWSYYMATLKNVTEVKGYTKGGPG HEEPIPMATYGWNLVKLHSGVDVFYKPTEQVDTLLCDIGESSSNPTIE EGRTLRVLKMVEPWLSSKPEFCIKVLNPYMPTVIEELEKLQRKHGGN LVRCPLSRNSTHEMYWVSGASGNIVSSVNTTSKMLLNRFTTRHRKPT YEKDVDLGAGTRSVSTETEKPDMTIIGRRLQRLQEEHKETWHYDQEN PYRTWAYHGSYEAPSTGSASSMVNGVVKLLTKPWDVIPMVTQLAMT DTTPFGQQRVFKEKVDTRTPQPKPGTRMVMTTTANWLWALLGKKK NPRLCTREEFISKVRSNAAIGAVFQEEQGWTSASEAVNDSRFWELVD KERALHQEGKCESCVYNMMGKREKKLGEFGRAKGSRAIWYMWLG ARFLEFEALGFLNEDHWFGRENSWSGVEGEGLHRLGYILEEIDKKDG DLMYADDTAGWDTRITEDDLQNEELITEQMAPHHKILAKAIFKLTYQ NKVVKVLRPTPRGAVMDIISRKDQRGSGQVGTYGLNTFTNMEVQLIR QMEAEGVITQDDMQNPKGLKERVEKWLKECGVDRLKRMAISGDDC VVKPLDERFGTSLLFLNDMGKVRKDIPQWEPSKGWKNWQEVPFCSH HFHKIFMKDGRSLVVPCRNQDELIGRARISQGAGWSLRETACLGKAY AQMWSLMYFHRRDLRLASMAICSAVPTEWFPTSRTTWSIHAHHQW MTTEDMLKVWNRVWIEDNPNMTDKTPVHSWEDIPYLGKREDLWCG SLIGLSSRATWAKNIHTAITQVRNLIGKEEYVDYMPVMKRYSAPSESE GVL Construct 1 MDAMKRGLCCVLLLCGAVFVSPFHLTTRNGEPHMIVSRQEKGKSLLF 263 KTENGVNMCTLMAMDLGELCEDTITYNCPLLRQNEPEDIDCWCNSTS TWVTYGTCTATGEHRREKRSVALVPHVGMGLETRTETWMSSEGAW KHAQRIETWVLRHPGFTIMAAILAYTIGTTYFQRVLIFILLTAVAPSMT MRCIGISNRDFVEGVSGGSWVDIVLEHGSCVTTMAKNKPTLDFELIKT EAKHPATLRKYCIEAKLTNTTTASRCPTQGEPSLNEEQDKRFVCKHS MVDRGWGNGCGLFGKGGIVTCAMFTCKKNMEGKIVQPENLEYTIVI TPHSGEEGNDTGKHGKEIKVTPQSSITEAELTGYGTVTMECSPRTGLD FNEMVLLQMENKAWLVHRQWFLDLPLPWLPGADTQGSNWIQKETL VTFKNPHAKKQDVVVLGSQEGAMHTALTGATEIQMSSGNLLFTGHL KCRLRMDKLQLKGMSYSMCTGKFKVVKEIAETQHGTIVIRVQYEGD GSPCKIPFEIMDLEKRHVLGRLITVNPIVTEKDSPVNIEAEPPFGDSYIII GVEPGQLKLSWFKKGSSIGQMFETTMRGAKRMAILGDTAWDFGSLG GVFTSIGKALHQVFGAIYGAAFSGVSWTMKILIGVVITWIGMNSRSTS LSVSLVLVGVVTLYLGVMVQA Construct 2 MDAMKRGLCCVLLLCGAVFVSPFHLTTRNGEPHMIVSRQEKGKSLLF 264 KTENGVNMCTLMAMDLGELCEDTITYNCPLLRQNEPEDIDCWCNSTS TWVTYGTCTATGEHRREKRSVALVPHVGMGLETRTETWMSSEGAW KHAQRIETWVLRHPGFTIMAAILAYTIGTTYFQRVLIFILLTAVAPSMT MRCIGISNRDFVEGVSGGSWVDIVLEHGSCVTTMAKNKPTLDFELIKT EAKHPATLRKYCIEAKLTNTTTASRCPTQGEPSLNEEQDKRFVCKHS MVDRGWGNGCGLFGKGGIVTCAMFTCKKNMEGKIVQPENLEYTIVI TPHSGEEGNDTGKHGKEIKVTPQSSITEAELTGYGTVTMECSPRTGLD FNEMVLLQMENKAWLVHRQWFLDLPLPWLPGADTQGSNWIQKETL VTFKNPHAKKQDVVVLGSQEGAMHTALTGATEIQMSSGNLLFTGHL KCRLRMDKLQLKGMSYSMCTGKFKVVKEIAETQHGTIVIRVQYEGD GSPCKIPFEIMDLEKRHVLGRLITVNPIVTEKDSPVNIEAEPPFGDSYIII GVEPGQLKLSWFKKG Construct 3 MDAMKRGLCCVLLLCGAVFVSPFHLTTRNGEPHMIVSRQEKGKSLLF 265 KTENGVNMCTLMAMDLGELCEDTITYNCPLLRQNEPEDIDCWCNSTS TWVTYGTCTATGEHRREKRSVALVPHVGMGLETRTETWMSSEGAW KHAQRIETWVLRHPGFTIMAAILAYTIGTTYFQRVLIFILLTAVAPSMT MRCIGISNRDFVEGVSGGSWVDIVLEHGSCVTTMAKNKPTLDFELIKT EAKHPATLRKYCIEAKLTNTTTASRCPTQGEPSLNEEQDKRFVCKHS MVDRGWGNGCGLFGKGGIVTCAMFTCKKNMEGKIVQPENLEYTIVI TPHSGEEGNDTGKHGKEIKVTPQSSITEAELTGYGTVTMECSPRTGLD FNEMVLLQMENKAWLVHRQWFLDLPLPWLPGADTQGSNWIQKETL VTFKNPHAKKQDVVVLGSQEGAMHTALTGATEIQMSSGNLLFTGHL KCRLRMDKLQLKGMSYSMCTGKFKVVKEIAETQHGTIVIRVQYEGD GSPCKIPFEIMDLEKRHVLGRLITVNPIVTEKDSPVNIEAEPPFGDSYIII GVEPGQLKLSWFKKGGGGGSGGGGSGGGGSEVKLQQSGTEVVKPG ASVKLSCKASGYIFTSYDIDWVRQTPEQGLEWIGWIFPGEGSTEYNEK FKGRATLSVDKSSSTAYMELTRLTSEDSAVYFCARGDYYRRYFDLW GQGTTVTVSSGGGGSGGGGSGGGGSDIQMTQSPSFLSTSLGNSITITC HASQNIKGWLAWYQQKSGNAPQLLIYKASSLQSGVPSRFSGSGSGTD YIFTISNLQPEDIATYYCQHYQSFPWTFGGGTKLEIKRDYKDDDDK Construct 4 MDWTWILFLVAAATRVHSKGMSYSMCTGKFKVVKEIAETQHGTIVI 266 RVQTEGDGSPCKIPFEIMDLEKRHVLGRLITVNPIVTEKDSPVNIEAEPP FGDSYIIIGVEPGQLKLNWFKKGSSIGQMFETTMRGAKRMAILSGGDII KLLNEQVNKEMQSSNLYMSMSSWCYTHSLDGAGLFLFDHAAEEYEH AKKLIIFLNENNVPVQLTSISAPEHKFEGLTQIFQKAYEHEQHISESINNI VDHAIKSKDHATFNFLQWYVAEQHEEEVLFKDILDKIELIGNENHGL YLADQYVKGIAKSRKS Dengue 2 prME MLNILNRRRRTAGIIIMMIPTVMA FHLTTRNGEPHMIVSRQEKGKSL 267 (Thailand/0168/ LFKTEDGVNMCTLMAMDLGELCEDTITYKCPLLRQNEPEDIDCWCNS 1979) TSTWVTYGTCTTTGEHRREKRSVALVPHVGMGLETRTETWMSSEGA WKHAQRIETWILRHPGFTIMAAILAYTIGTTHFQRALIFILLTAVAPSM TMRCIGISNRDFVEGVSGGSWVDIVLEHGSCVTTMAKNKPTLDFELIK TEAKQPATLRKYCIEAKLTNTTTESRCPTQGEPSLNEEQDKRFVCKHS MVDRGWGNGCGLFGKGGIVTCAMFTCKKNMEGKIVQPENLEYTIVV TPHSGEEHAVGNDTGKHGKEIKVTPQSSITEAELTGYGTVTMECSPRT GLDFNEMVLLQMENKAWLVHRQWFLDLPLPWLPGADTQGSNWIQK ETLVTFKNPHAKKQDVVVLGSQEGAMHTALTGATEIQMSSGNLLFT GHLKCRLRMDKLQLKGMSYSMCTGKFKVVKEIAETQHGTIVIRVQY EGDGSPCKIPFEIMDLEKRHVLGRLITVNPIVTEKDSPVNIEAEPPFGDS YIIIGVEPGQLKLNWFKKGSSIGQMFETTMRGAKRMAILGDTAWDFG SLGGVFTSIGKALHQVFGAIYGAAFSGVSWTMKILIGVIITWIGMNSRS TSLSVSLVLVGIVTLYLGVMVQA Dengue 2 prME MLNILNRRRRTAGIIIMMIPTVMA FHLTTRNGEPHMIVGRQEKGKS 268 (Thailand/16681/ LLFKTEDGVNMCTLMAIDLGELCEDTITYKCPLLRQNEPEDIDCWCNS 1984) TSTWVTYGTCATTGEHRREKRSVALVPHVGMGLETRTETWMSSEGA WKHVQRIETWILRHPGFTIMAAILAYTIGTTHFQRALIFILLTAVAPSM TMRCIGMSNRDFVEGVSGGSWVDIVLEHGSCVTTMAKNKPTLDFELI KTEAKQPATLRKYCIEAKLTNTTTESRCPTQGEPSLNEEQDKRFVCKH SMVDRGWGNGCGLFGKGGIVTCAMFRCKKNMEGKVVQPENLEYTI VITPHSGEEHAVGNDTGKHGKEIKITPQSSTTEAELTGYGTVTMECSP RTGLDFNEMVLLQMENKAWLVHRQWFLDLPLPWLPGADTQGSNWI QKETLVTFKNPHAKKQDVVVLGSQEGAMHTALTGATEIQMSSGNLL FTGHLKCRLRMDKLQLKGMSYSMCTGKFKVVKEIAETQHGTIVIRVQ YEGDGSPCKIPFEIMDLEKRHVLGRLITVNPIVTEKDSPVNIEAEPPFGD SYIIIGVEPGQLKLNWFKKGSSIGQMFETTMRGAKRMAILGDTAWDF GSLGGVFTSIGKALHQVFGAIYGAAFSGVSWTMKILIGVIITWIGMNS RSTSLSVTLVLVGIVTLYLGVMVQA Dengue 2 prME MLNILNRRRRTAGIIIMMIPTVMA FHLTTRNGEPHMIVGRQEKGKS 269 (Jamaica/1409/ LLFKTEDGVNMCTLMAIDLGELCEDTITYKCPLLRQNEPEDIDCWCNS 1983) TSTWVTYGTCATTGEHRREKRSVALVPHVGMGLETRTETWMSSEGA WKHVQRIETWILRHPGFTIMAAILAYTIGTTHFQRALIFILLTAVAPSM TMRCIGISNRDFVEGVSGGSWVDIVLEHGSCVTTMAKNKPTLDFELIK TEAKQPATLRKYCIEAKLTNTTTESRCPTQGEPSLNEEQDKRFLCKHS MVDRGWGNGCGLFGKGGIVTCAMFTCKKNMEGKVVLPENLEYTIVI TPHSGEEHAVGNDTGKHGKEIKITPQSSITEAELTGYGTVTMECSPRT GLDFNEMVLLQMEDKAWLVHRQWFLDLPLPWLPGADTQGSNWIQK ETLVTFKNPHAKKQDVVVLGSQEGAMHTALTGATEIQMSSGNLLFT GHLKCRLRMDKLQLKGMSYSMCTGKFKIVKEIAETQHGTIVIRVQYE GDGSPCKIPFEIMDLEKRHVLGRLITVNPIVTEKDSPVNIEAEPPFGDSY IIIGVEPGQLKLNWFKKGSSIGQMFETTMRGAKRMAILGDTAWDFGS LGGVFTSIGKALHQVFGAIYGAAFSGVSWTMKILIGVIITWIGMNSRST SLSVSLVLVGVVTLYLGAMVQA Dengue 2 prME MLNILNRRRRTAGIIIMMIPTVMA FHLTTRNGEPHMIVSRQEKGKSL 270 (Thailand/NGS- LFKTEDGVNMCTLMAMDLGELCEDTITYKCPFLKQNEPEDIDCWCNS C/1944) TSTWVTYGTCTTTGEHRREKRSVALVPHVGMGLETRTETWMSSEGA WKHAQRIETWILRHPGFTIMAAILAYTIGTTHFQRALIFILLTAVAPSM TMRCIGISNRDFVEGVSGGSWVDIVLEHGSCVTTMAKNKPTLDFELIE TEAKQPATLRKYCIEAKLTNTTTDSRCPTQGEPSLNEEQDKRFVCKHS MVDRGWGNGCGLFGKGGIVTCAMFTCKKNMKGKVVQPENLEYTIVI TPHSGEEHAVGNDTGKHGKEIKITPQSSITEAELTGYGTVTMECSPRT GLDFNEMVLLQMENKAWLVHRQWFLDLPLPWLPGADTQGSNWIQK ETLVTFKNPHAKKQDVVVLGSQEGAMHTALTGATEIQMSSGNLLFT GHLKCRLRMDKLQLKGMSYSMCTGKFKVVKEIAETQHGTIVIRVQY EGDGSPCKIPFEIMDLEKRHVLGRLITVNPIVTEKDSPVNIEAEPPFGDS YIIIGVEPGQLKLNWFKKGSSIGQMIETTMRGAKRMAILGDTAWDFGS LGGVFTSIGKALHQVFGAIYGAAFSGVSWIMKILIGVIITWIGMNSRST SLSVSLVLVGVVTLYLGVMVQA Dengue 2 prME MLNILNRRRRTAGIIIMMIPTVMA FHLTTRNGEPHMIVSRQEKGKSL 271 (PuertoRico/ LFKTKDGTNMCTLMAMDLGELCEDTITYKCPFLKQNEPEDIDCWCNS PR159-S1/1969) TSTWVTYGTCTTTGEHRREKRSVALVPHVGMGLETRTETWMSSEGA WKHAQRIETWILRHPGFTIMAAILAYTIGTTHFQRVLIFILLTAIAPSMT MRCIGISNRDFVEGVSGGSWVDIVLEHGSCVTTMAKNKPTLDFELIKT EAKQPATLRKYCIEAKLTNTTTDSRCPTQGEPTLNEEQDKRFVCKHS MVDRGWGNGCGLFGKGGIVTCAMFTCKKNMEGKIVQPENLEYTVVI TPHSGEEHAVGNDTGKHGKEVKITPQSSITEAELTGYGTVTMECSPRT GLDFNEMVLLQMKDKAWLVHRQWFLDLPLPWLPGADTQGSNWIQK ETLVTFKNPHAKKQDVVVLGSQEGAMHTALTGATEIQMSSGNLLFT GHLKCRLRMDKLQLKGMSYSMCTGKFKVVKEIAETQHGTIVIRVQY EGDGSPCKTPFEIMDLEKRHVLGRLTTVNPIVTEKDSPVNIEAEPPFGD SYIIIGVEPGQLKLDWFKKGSSIGQMFETTMRGAKRMAILGDTAWDF GSLGGVFTSIGKALHQVFGAIYGAAFSGVSWTMKILIGVIITWIGMNS RSTSLSVSLVLVGIVTLYLGVMVQA Dengue 2 prME MLNILNRRRRTAGIIIMMIPTVMA FHLTTRNGEPHMIVSRQEKGKSL 272 (16681-PDK53) LFKTEVGVNMCTLMAMDLGELCEDTITYKCPLLRQNEPEDIDCWCNS TSTWVTYGTCTTMGEHRREKRSVALVPHVGMGLETRTETWMSSEGA WKHVQRIETWILRHPGFTMMAAILAYTIGTTHFQRALILILLTAVTPS MTMRCIGMSNRDFVEGVSGGSWVDIVLEHGSCVTTMAKNKPTLDFE LIKTEAKQPATLRKYCIEAKLTNTTTESRCPTQGEPSLNEEQDKRFVC KHSMVDRGWGNGCGLFGKGGIVTCAMFRCKKNMEGKVVQPENLEY TIVITPHSGEEHAVGNDTGKHGKEIKITPQSSITEAELTGYGTITMECSP RTGLDFNEIVLLQMENKAWLVHRQWFLDLPLPWLPGADTQGSNWIQ KETLVTFKNPHAKKQDVVVLGSQEGAMHTALTGATEIQMSSGNLLF TGHLKCRLRMDKLQLKGMSYSMCTGKFKVVKEIAETQHGTIVIRVQ YEGDGSPCKIPFEIMDLEKRHVLGRLITVNPIVTEKDSPVNIEAEPPFGD SYIIIGVEPGQLKLNWFKKGSSIGQMFETTMRGAKRMAILGDTAWDF GSLGGVFTSIGKALHQVFGAIYGAAFSGVSWTMKILIGVIITWIGMNS RSTSLSVTLVLVGIVTLYLGVMVQA Dengue 2 prME MLNILNRRRRTAGIIIMMIPTVMA FHLTTRNGEPHMIVSRQEKGKSL 273 (Peru/IQT2913/ LFKTKDGTNMCTLMAMDLGELCEDTITYKCPFLKQNEPEDIDCWCNS 1996) TSTWVTYGTCTTTGEHRREKRSVALVPHVGMGLETRTETWMSSEGA WKHAQRIETWILRHPGFTIMAAILAYTIGTTHFQRVLIFILLTAIAPSMT MRCIGISNRDFVEGVSGGSWVDIVLEHGSCVTTMAKNKPTLDFELIKT EAKQPATLRKYCIEAKLTNTTTDSRCPTQGEPTLNEEQDKRFVCKHS MVDRGWGNGCGLFGKGGIVTCAMFTCKKNMEGKIVQPENLEYTVVI TPHSGEEHAVGNDTGKHGKEVKITPQSSITEAELTGYGTVTMECSPRT GLDFNEMVLLQMEDKAWLVHRQWFLDLPLPWLPGADTQGSNWIQK ETLVTFKNPHAKKQDVVVLGSQEGAMHTALTGATEIQMSSGNLLFT GHLKCRLRMDKLQLKGMSYSMCTGKFKIVKEIAETQHGTIVIRVQYE GDGSPCKIPFEIMDLEKRHVLGRLITVNPIVTEKDSPVNIEAEPPFGDSY IIIGAEPGQLKLDWFKKGSSIGQMFETTMRGAKRMAILGDTAWDFGS LGGVFTSIGKALHQVFGAIYGAAFSGVSWTMKILIGVIITWIGMNSRST SLSVSLVLVGIVTLYLGVMVQA Dengue 2 prME MLNILNRRRRTAGIIIMMIPTVMA FHLTTRNGEPHMIVSRQEKGKSL 274 (Thailand/PUO- LFKTEDGVNMCTLMAMDLGELCEDTITYKCPLLRQNEPEDIDCWCNS 218/1980) TSTWVTYGTCTTTGEHRREKRSVALVPHVGMGLETRTETWMSSEGA WKHAQRIEIWILRHPGFTIMAAILAYTIGTTHFQRALIFILLTAVAPSMT MRCIGISNRDFVEGVSGGSWVDIVLEHGSCVTTMAKNKPTLDFELIKT EAKQPATLRKYCIEAKLTNTTTESRCPTQGEPSLNEEQDKRFVCKHSM VDRGWGNGCGLFGKGGIVTCAMFTCKKNMEGKVVQPENLEYTIVVT PHSGEEHAVGNDTGKHGKEIKVTPQSSITEAELTGYGTVTMECSPRTG LDFNEMVLLQMENKAWLVHRQWFLDLPLPWLPGADTQGSNWIQKE TLVTFKNPHAKKQDVVVLGSQEGAMHTALTGATEIQMSSGNLLFTG HLKCRLRMDKLQLKGMSYSMCTGKFKVVKEIAETQHGTIVIRVQYE GDGSPCKIPFEIMDLEKRHVLGRLITVNPIVTEKDSPVNIEAEPPFGDSY IIIGVEPGQLKLNWFKKGSSIGQMFETTMRGAKRMAILGDTAWDFGS LGGVFTSIGKALHQVFGAIYGAAFSGVSWTMKILIGVIITWIGMNSRST SLSVSLVLVGIVTLYLGVMVQA Dengue 2 prME MLNILNRRRRTAGIIIMMIPTVMA FHLTTRNGEPHMIVSRQEKGKSL 275 (D2Y98P) with LFKTENGVNMCTLMAMDLGELCEDTITYNCPLLRQNEPEDIDCWCNS native leader TSTWVTYGTCTATGEHRREKRSVALVPHVGMGLETRTETWMSSEGA WKHAQRIETWVLRHPGFTIMAAILAYTIGTTYFQRVLIFILLTAVAPS MTMRCIGISNRDFVEGVSGGSWVDIVLEHGSCVTTMAKNKPTLDFEL IKTEAKHPATLRKYCIEAKLTNTTTASRCPTQGEPSLNEEQDKRFVCK HSMVDRGWGNGCGLFGKGGIVTCAMFTCKKNMEGKIVQPENLEYTI VITPHSGEENAVGNDTGKHGKEIKVTPQSSITEAELTGYGTVTMECSP RTGLDFNEMVLLQMENKAWLVHRQWFLDLPLPWLPGADTQGSNWI QKETLVTFKNPHAKKQDVVVLGSQEGAMHTALTGATEIQMSSGNLL FTGHLKCRLRMDKLQLKGMSYSMCTGKFKVVKEIAETQHGTIVIRVQ YEGDGSPCKIPFEIMDLEKRHVLGRLITVNPIVTEKDSPVNIEAEPPFGD SYIIIGVEPGQLKLSWFKKGSSIGQMFETTMRGAKRMAILGDTAWDF GSLGGVFTSIGKALHQVFGAIYGAAFSGVSWTMKILIGVVITWIGMNS RSTSLSVSLVLVGVVTLYLGVMVQA gi|158348409|ref| MNNQRKKTGRPSFNMLKRARNRVSTVSQLAKRFSKGLLSGQGPMKL 276 NP_722466.2| capsid VMAFIAFLRFLAIPPTAGILARWGSFKKNGAIKVLRGFKKEISNMLNI protein [Dengue MNRRKR virus 1] gi|164654862|ref| MNNQRKKTGKPSINMLKRVRNRVSTGSQLAKRFSKGLLNGQGPMKL 277 YP_001531164.2| VMAFIAFLRFLAIPPTAGVLARWGTFKKSGAIKVLKGFKKEISNMLSII Capsid protein NQRKK [Dengue virus 3] gi|159024809|ref| MNNQRKKAKNTPFNMLKRERNRVSTVQQLTKRFSLGMLQGRGPLKL 278 NP_739591.2| FMALVAFLRFLTIPPTAGILKRWGTIKKSKAINVLRGFRKEIGRMLNIL Capsid protein NRRRR [Dengue virus 2] gi|158348408|ref| MNNQRKKTGRPSFNMLKRARNRVSTVSQLAKRFSKGLLSGQGPMKL 279 NP_722457.2| VMAFIAFLRFLAIPPTAGILARWGSFKKNGAIKVLRGFKKEISNMLNI anchored capsid MNRRKRSVTMLLMLLPTALA protein [Dengue virus 1] gi|164654854|ref| MNNQRKKTGKPSINMLKRVRNRVSTGSQLAKRFSKGLLNGQGPMKL 280 YP_001531165.2| VMAFIAFLRFLAIPPTAGVLARWGTFKKSGAIKVLKGFKKEISNMLSII Anchored capsid NQRKKTSLCLMMILPAALA protein [Dengue virus 3] gi|159024808|ref| MNNQRKKAKNTPFNMLKRERNRVSTVQQLTKRFSLGMLQGRGPLKL 281 NP_739581.21 FMALVAFLRFLTIPPTAGILKRWGTIKKSKAINVLRGFRKEIGRMLNIL Anchored capsid NRRRRSAGMIIMLIPTVMA protein [Dengue virus 2] gi|73671168|ref| MNQRKKVVRPPFNMLKRERNRVSTPQGLVKRFSTGLFSGKGPLRMV 282 NP_740314.1| LAFITFLRVLSIPPTAGILKRWGQLKKNKAIKILIGFRKEIGRMLNILNG anchored capsid RKRSTITLLCLIPTVMA (anchC) protein [Dengue virus 4] gi|73671167|ref| MNQRKKVVRPPFNMLKRERNRVSTPQGLVKRFSTGLFSGKGPLRMV 283 NP_740313.1| virion LAFITFLRVLSIPPTAGILKRWGQLKKNKAIKILIGFRKEIGRMLNILNG capsid (virC) RKR protein [Dengue virus 4] gi|164654853|ref| MRCVGVGNRDFVEGLSGATWVDVVLEHGGCVTTMAKNKPTLDIEL 284 YP_001531168.2| QKTEATQLATLRKLCIEGKITNITTDSRCPTQGEAVLPEEQDQNYVCK Envelope protein HTYVDRGWGNGCGLFGKGSLVTCAKFQCLEPIEGKVVQYENLKYTV [Dengue virus 3] IITVHTGDQHQVGNETQGVTAEITPQASTTEAILPEYGTLGLECSPRTG LDFNEMILLTMKNKAWMVHRQWFFDLPLPWASGATTETPTWNRKE LLVTFKNAHAKKQEVVLGSQEGAMHTALTGATEIQNSGGTSIFAGH LKCRLKMDKLELKGMSYAMCTNTFVLKKEVSETQHGTILIKVEYKG EDAPCKIPFSTEDGQGKAHNGRLITANPVVTKKEEPVNIEAEPPFGESN IVIGIGDNALKINWYKKGSSIGKMFEATERGARRMAILGDTAWDFGS VGGVLNSLGKMVHQIFGSAYTALFSGVSWVMKIGIGVLLTWIGLNSK NTSMSFSCIAIGIITLYLGAVVQA gi|158828123|ref| MRCVGIGNRDFVEGLSGATWVDVVLEHGSCVTTMAKDKPTLDIELL 285 NP_722460.2| KTEVTNPAVLRKLCIEAKISNTTTDSRCPTQGEATLVEEQDTNFVCRR envelope protein TFVDRGWGNGCGLFGKGSLITCAKFKCVTKLEGKIVQYENLKYSVIV [Dengue virus 1] TVHTGDQHQVGNETTEHGTTATITPQAPTSEIQLTDYGALTLDCSPRT GLDFNEMVLLTMKKKSWLVHKQWFLDLPLPWTSGASTSQETWNRQ DLLVTFKTAHAKKQEVVVLGSQEGAMHTALTGATEIQTSGTTTIFAG HLKCRLKMDKLILKGMSYVMCTGSFKLEKEVAETQHGTVLVQVKYE GTDAPCKIPFSSQDEKGVTQNGRLITANPIVTDKEKPVNIEAEPPFGES YIVVGAGEKALKLSWFKKGSSIGKMFEATARGARRMAILGDTAWDF GSIGGVFTSVGKLIHQIFGTAYGVLFSGVSWTMKIGIGILLTWLGLNSR STSLSMTCIAVGMVTLYLGVMVQA gi|159024812|ref| MRCIGMSNRDFVEGVSGGSWVDIVLEHGSCVTTMAKNKPTLDFELIK 286 NP_739583.2| TEAKQPATLRKYCIEAKLTNTTTESRCPTQGEPSLNEEQDKRFVCKHS Envelope protein MVDRGWGNGCGLFGKGGIVTCAMFRCKKNMEGKVVQPENLEYTIVI [Dengue virus 2] TPHSGEEHAVGNDTGKHGKEIKITPQSSITEAELTGYGTVTMECSPRT GLDFNEMVLLQMENKAWLVHRQWFLDLPLPWLPGADTQGSNWIQK ETLVTFKNPHAKKQDVVVLGSQEGAMHTALTGATEIQMSSGNLLFT GHLKCRLRMDKLQLKGMSYSMCTGKFKVVKEIAETQHGTIVIRVQY EGDGSPCKIPFEIMDLEKRHVLGRLITVNPIVTEKDSPVNIEAEPPFGDS YIIIGVEPGQLKLNWFKKGSSIGQMFETTMRGAKRMAILGDTAWDFG SLGGVFTSIGKALHQVFGAIYGAAFSGVSWTMKILIGVIITWIGMNS RS TSLSVTLVLVGIVTLYLGVMVQA tr|Q9IZI6|Q9IZI6_9 MRCVGVGNRDFVEGVSGGAWVDLVLEHGGCVTTMAQGKPTLDFEL 287 FLAV Envelope TKTTAKEVALLRTYCIEASISNITTATRCPTQGEPYLKEEQDQQYICRR protein (Fragment) DVVDRGWGNGCGLFGKGGVVTCAKFSCSGKITGNLVRIENLEYTVV OS = Dengue virus 4 VTVHNGDTHAVGNDTSNHGVTAMITPRSPSVEVKLPDYGELTLDCEP GN = E PE = 4 RSGIDFNEMILMKMKKKTWLVHKQWFLDLPLPWTAGADTSEVHWN SV = 1 YKERMVTFKVPHAKRQDVTVLGSQEGAMHSALAGATEVDSGDGNH MFAGHLKCEVRMEKLRIKGMSYTMCSGKFSIDKEMAETQHGTTVVK VKYEGAGAPCKVPIEIRDVNKEKVVGRIISSTPLAENTNSVTNIELEPPF GDSYIVIGVGNSALTLHWFRKGSSIGKMFESTYRGAKRMAILGETAW DFGSVGGLFTSLGKAVHQVFGSVYTTMFGGVSWMIRILIGFLVLWIG TNSRNTSMAMTCIAVGGITLFLGF gi|73671170|ref| SVALTPHSGMGLETRAETWMSSEGAWKHAQRVESWILRNPGFALLA 288 NP_740316.11 GFMAYMIGQTGIQRTVFFVLMMLVAPSYG membrane (M) protein [Dengue virus 4] gi|158828127|ref| SVALAPHVGMGLDTRTQTWMSAEGAWRQVEKVETWALRHPGFTIL 289 YP_001531167.1| ALFLAHYIGTSLTQKVVIFILLMLVTPSMT Membrane glycoprotein [Dengue virus 3] gi|158828122|ref| SVALAPHVGLGLETRTETWMSSEGAWKQIQKVETWALRHPGFTVIA 290 NP_722459.2| LFLAHAIGTSITQKGIIFILLMLVTPSMA membrane glycoprotein [Dengue virus 1] gi|159024811|ref| SVALVPHVGMGLETRTETWMSSEGAWKHVQRIETWILRHPGFTMMA 291 NP_739592.2| AILAYTIGTTHFQRALIFILLTAVTPSMT Membrane glycoprotein [Dengue virus 2]

The first underlined sequence corresponds to a signal peptide, which may be omitted from each sequence. Thus, any RNA vaccine provided herein may encode an antigen represented by a sequence of Table 29, with or without the underlined signal peptide.

TABLE 30 Dengue Antigen polynucleotides Construct Gene Number ID Description Construct 1 131502 Dengue 2, D2Y98P strain, PrME DEN2_D2Y98P_PrME_Hs3 transmembrane antigen 2 131503 Dengue 2, D2Y98P strain, PrME secreted DEN2_D2Y98P_PrME80_Hs3 antigen 3 131507 Dengue 2, D2Y98P strain, PrME secreted DEN2_D2Y98P_PrME80_ScFv.aDEC205.FLAG_Hs3 antigen with dendritic targeting ScFv against mouse DEC205 4 120554 Dengue strain 2 domain 3 ferritin DEN2_DIII_Ferritin_Hs3

TABLE 31 Detailed experimental design (treatment, readouts) Mouse Formulation/ Group Strain Vaccine* Chemistry Route Dose Readouts 1 Female N/A N/A N/A Serum samples 2 BALB/c DEN2Y98- N1- ID 0.4 mg/kg collected on weeks 3 6-8 weeks PrME methylpseudouridine/ IM in 1, 3, and 5. of age (construct 1 5- LNP Serum analyzed via 4 from Tables methyl ID 0.08 mg/kg Western blot 5 30) cytosine IM in LNP 6 ID 0.016 mg/kg 7 IM in LNP 8 N1- ID 0.4 mg/kg 9 methylpseudouridine IM in LNP 10 ID 0.08 mg/kg 11 IM in LNP 11 ID 0.016 mg/kg 12 IM in LNP 13 DEN2Y98- N1- ID 0.4 mg/kg 14 PrME80 methylpseudouridine/ IM in (construct 2 5- LNP 15 from Table methyl ID 0.08 mg/kg 16 30) cytosine IM in LNP 17 ID 0.016 mg/kg 18 IM in LNP 19 N1- ID 0.4 mg/kg 20 methylpseudouridine IM in LNP 21 ID 0.08 mg/kg 22 IM in LNP 23 ID 0.016 mg/kg 24 IM in LNP 25 DEN2Y98- N1- ID 0.4 mg/kg 26 PrME80-DC methylpseudouridine/ IM in (construct 3 5- LNP 27 from Table methyl ID 0.08 mg/kg 28 30) cytosine IM in LNP 29 ID 0.016 mg/kg 30 IM in LNP 31 N1- ID 0.4 mg/kg 32 methylpseudouridine IM in LNP 33 ID 0.08 mg/kg 34 IM in LNP 35 ID 0.016 mg/kg 36 IM in LNP 37 DEN2-DIII- N1- ID 0.4 mg/kg 38 Ferritin methylpseudouridine IM in LNP 39 (construct 4 ID 0.08 mg/kg 40 from Table 30) IM in LNP 41 ID 0.016 mg/kg 42 IM in LNP 43 Control, — IV 10⁵ PFU D2Y98P live virus *(n = 10, female) mice/group) Delivered week 0 and 3

TABLE 32 Example 30 Alleles Tested Allele A*01:01 A*02:01 A*03:01 A*11:01 A*24:02 B*07:02 B*27:05 H-2Kb

TABLE 33 ProImmune REVEAL ® binding assay data for A*01:01 Peptide I.D. SEQ ID NO REVEAL ® score at 0 h TTDISEMGA 436 68.4

TABLE 34 ProImmune REVEAL ® binding assay data for A*02:01 Peptide I.D. SEQ ID NO REVEAL ® score at 0 h TMWHVTRGA 437 112.0 MWHVTRGAV 438  62.7 GLYGNGVVT 439  87.7 TLILAPTRV 440 104.2 LILAPTRVV 441 106.4 ILAPTRVVA 442  95.7 VVAAEMEEA 443  92.2 IVDLMCHAT 444  62.7 LMCHATFTM 445  72.9 MGEAAAIFM 446  50.6 GEAAAIFMT 447  74.3 KTVWFVPSI 448 115.9 LMRRGDLPV 449  82.3 TLLCDIGES 450  63.9 LLCDIGESS 451  93.9 AMTDTTPFG 452  91.9 GQQRVFKEK 453  47.1 KLTYQNKVV 454  92.3 AISGDDCVV 455  91.1 LMYFHRRDL 456  97.8

TABLE 35 ProImmune REVEAL ® binding assay data for A*03:01 Peptide I.D. SEQ ID NO REVEAL ® score at 0 h RTLILAPTR 457 91.4 TLILAPTRV 458 55.2 MCHATFTMR 459 86.8 TVWFVPSIK 460 53.6 GQQRVFKEK 461 59.6 CVYNMMGKR 462 81.6

TABLE 36 ProImmune REVEAL ® binding assay data for A*11:01 Peptide I.D. SEQ ID NO REVEAL ® score at 0 h HTMWHVTRG 463  56.3 RTLILAPTR 464  89.9 TLILAPTRV 465  59.0 MCHATFTMR 466  91.0 ATFTMRLLS 467  58.5 GEAAAIFMT 468  50.3 KTVWFVPSI 292  50.8 TVWFVPSIK 293  92.2 GQQRVFKEK 294  85.5 CVYNMMGKR 295 113.2 VYNMMGKRE 296  62.5 YNMMGKREK 297  80.9 NMMGKREKK 298  77.9 GTYGLNTFT 299  63.6 ISGDDCVVK 300  88.7

TABLE 37 ProImmune REVEAL ® binding assay data for A*24:02 Peptide I.D. SEQ ID NO REVEAL ® score at 0 h LMCHATFTM 301  99.5 CHATFTMRL 302  75.9 GEAAAIFMT 303  58.9 KTVWFVPSI 304  89.1 HWTEAKMLL 305 103.2 WTEAKMLLD 306  94.7 LGCGRGGWS 307  74.8 MAMTDTTPF 308  51.3 MYADDTAGW 309  76.8 VGTYGLNTF 310  96.0 YFHRRDLRL 311  87.5

TABLE 38 ProImmune REVEAL ® binding assay data for B*07:02 Peptide I.D. SEQ ID NO REVEAL ® score at 0 h FKPGTSGSP 312  50.4 KPGTSGSPI 313 112.1 IPERSWNSG 314  45.2 PERVILAGP 315  56.1 LMRRGDLPV 316 178.9 PLSRNSTHE 317  65.0 LSRNSTHEM 318 124.5 SRNSTHEMY 319  52.0 MAMTDTTPF 320 117.4 TPFGQQRVF 321 112.7 LMYFHRRDL 322 119.6

TABLE 39 ProImmune REVEAL ® binding assay data for B*27:05 Peptide I.D. SEQ ID NO REVEAL ® score at 0 h LRTLILAPT 323  58.7 LMCHATFTM 324  98.2 ARGYISTRV 325 125.3 RRGDLPVWL 326 144.8 GQQRVFKEK 327  95.4 SRAIWYMWL 328  53.9 FKLTYQNKV 329  53.7

TABLE 40 ProImmune REVEAL ® binding assay data for H-2Kb Peptide I.D. SEQ ID NO REVEAL ® score at 0 h FKPGTSGSP 330  45.7 LAPTRVVAA 331 102.5 LMCHATFTM 332  59.0 CHATFTMRL 333  60.3 HATFTMRLL 334  69.5 ATFTMRLLS 335  55.6 KTVWFVPSI 336  54.4 LSRNSTHEM 337  51.1 QQRVFKEKV 338  63.4 YGLNTFTNM 339  75.4 LMYFHRRDL 340  54.9

TABLE 41 Exampe 30 Results A*02:01 Peptide ID REVEAL ® Score 5. KQWFLDLPL (SEQ ID NO: 341) 86.0 6. RQWFLDLPL (SEQ ID NO: 342) 77.7 7. RQWFFDLPL (SEQ ID NO: 343) 80.5 8. TALTGATEI (SEQ ID NO: 344)  0.9 Positive Control 100.0 +/- 4.8

TABLE 42 Full-length Dengue Amino Acid Sequences (Homo sapiens strains; Brazil, Cuba and U.S.) GenBank Collection Accession Length Type Country Genome Region Date Virus Name AGN94866 3392 1 Brazil UTR5CMENS1NS2A 2010 Dengue virus 1 isolate NS2BNS3NS4A2KNS 12898/BR-PE/10, 4BNS5UTR3 complete genome AGN94867 3392 1 Brazil UTR5CMENS1NS2A 2010 Dengue virus 1 isolate NS2BNS3NS4A2KNS 13501/BR-PE/10, 4BNS5UTR3 complete genome AGN94868 3392 1 Brazil UTR5CMENS1NS2A 2010 Dengue virus 1 isolate NS2BNS3NS4A2KNS 13671/BR-PE/10, 4BNS5UTR3 complete genome AGN94869 3392 1 Brazil UTR5CMENS1NS2A 2010 Dengue virus 1 isolate NS2BNS3NS4A2KNS 13861/BR-PE/10, 4BNS5UTR3 complete genome AGN94870 3392 1 Brazil UTR5CMENS1NS2A 2010 Dengue virus 1 isolate NS2BNS3NS4A2KNS 14985/BR-PE/10, 4BNS5UTR3 complete genome AGN94871 3392 1 Brazil UTR5CMENS1NS2A 1996 Dengue virus 1 isolate NS2BNS3NS4A2KNS 21814/BR-PE/96, 4BNS5UTR3 complete genome AGN94872 3392 1 Brazil UTR5CMENS1NS2A 1997 Dengue virus 1 isolate NS2BNS3NS4A2KNS 40604/BR-PE/97, 4BNS5UTR3 complete genome AGN94873 3392 1 Brazil UTR5CMENS1NS2A 1997 Dengue virus 1 isolate NS2BNS3NS4A2KNS 41111/BR-PE/97, 4BNS5UTR3 complete genome AGN94874 3392 1 Brazil UTR5CMENS1NS2A 1998 Dengue virus 1 isolate NS2BNS3NS4A2KNS 52082/BR-PE/98, 4BNS5UTR3 complete genome AGN94875 3392 1 Brazil UTR5CMENS1NS2A 1999 Dengue virus 1 isolate NS2BNS3NS4A2KNS 59049/BR-PE/99, 4BNS5UTR3 complete genome AGN94876 3392 1 Brazil UTR5CMENS1NS2A 2000 Dengue virus 1 isolate NS2BNS3NS4A2KNS 70523/BR-PE/00, 4BNS5UTR3 complete genome AGN94877 3392 1 Brazil UTR5CMENS1NS2A 2001 Dengue virus 1 isolate NS2BNS3NS4A2KNS 74488/BR-PE/01, 4BNS5UTR3 complete genome AGN94878 3392 1 Brazil UTR5CMENS1NS2A 2001 Dengue virus 1 isolate NS2BNS3NS4A2KNS 75861/BR-PE/01, 4BNS5UTR3 complete genome AGN94879 3392 1 Brazil UTR5CMENS1NS2A 2002 Dengue virus 1 isolate NS2BNS3NS4A2KNS 88463/BR-PE/02, 4BNS5UTR3 complete genome AGN94865 3392 1 Brazil UTR5CMENS1NS2A 2010 Dengue virus 1 isolate NS2BNS3NS4A2KNS 9808/BR-PE/10, 4BNS5UTR3 complete genome ACO06150 3392 1 Brazil UTR5CMENS1NS2A 2000 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/BR/BID- 4BNS5UTR3 V2374/2000, complete genome ACO06151 3392 1 Brazil UTR5CMENS1NS2A 2000 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/BR/BID- 4BNS5UTR3 V2375/2000, complete genome ACO06153 3392 1 Brazil UTR5CMENS1NS2A 2001 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/BR/BID- 4BNS5UTR3 V2378/2001, complete genome ACO06155 3392 1 Brazil UTR5CMENS1NS2A 2002 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/BR/BID- 4BNS5UTR3 V2381/2002, complete genome ACO06157 3392 1 Brazil UTR5CMENS1NS2A 2003 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/BR/BID- 4BNS5UTR3 V2384/2003, complete genome ACO06161 3392 1 Brazil UTR5CMENS1NS2A 2004 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/BR/BID- 4BNS5UTR3 V2389/2004, complete genome ACO06164 3392 1 Brazil UTR5CMENS1NS2A 2005 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/BR/BID- 4BNS5UTR3 V2392/2005, complete genome ACO06167 3392 1 Brazil UTR5CMENS1NS2A 2006 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/BR/BID- 4BNS5UTR3 V2395/2006, complete genome ACO06170 3392 1 Brazil UTR5CMENS1NS2A 2007 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/BR/BID- 4BNS5UTR3 V2398/2007, complete genome ACO06173 3392 1 Brazil UTR5CMENS1NS2A 2008 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/BR/BID- 4BNS5UTR3 V2401/2008, complete genome ACY70762 3392 1 Brazil UTR5CMENS1NS2A 2008 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/BR/BID- 4BNS5 V3490/2008, complete genome ACJ12617 3392 1 Brazil UTR5CMENS1NS2A Dengue virus 1 isolate NS2BNS3NS4A2KNS DF01-HUB01021093, 4BNS5UTR3 complete genome AHC08447 3392 1 Brazil CMENS1NS2ANS2B 2011 Dengue virus 1 strain NS3NS4A2KNS4BNS5 1266/2011/BR/RJ/2011 polyprotein gene, partial cds AHC08446 3392 1 Brazil CMENS1NS2ANS2B 2010 Dengue virus 1 strain NS3NS4A2KNS4BNS5 242/2010/BR/RJ/2010 polyprotein gene, partial cds AHC08448 3392 1 Brazil CMENS1NS2ANS2B 1988 Dengue virus 1 strain NS3NS4A2KNS4BNS5 36034/BR/RJ/1988 polyprotein gene, partial cds AHC08449 3392 1 Brazil CMENS1NS2ANS2B 1989 Dengue virus 1 strain NS3NS4A2KNS4BNS5 38159/BR/RJ/1989 polyprotein gene, partial cds AHC08450 3392 1 Brazil CMENS1NS2ANS2B 2000 Dengue virus 1 strain NS3NS4A2KNS4BNS5 66694/BR/ES/2000 polyprotein gene, partial cds AHC08451 3392 1 Brazil CMENS1NS2ANS2B 2001 Dengue virus 1 strain NS3NS4A2KNS4BNS5 68826/BR/RJ/2001 polyprotein gene, partial cds AGN94880 3391 2 Brazil UTR5CMENS1NS2A 2010 Dengue virus 2 isolate NS2BNS3NS4A2KNS 13858/BR-PE/10, 4BNS5UTR3 complete genome AGN94881 3391 2 Brazil UTR5CMENS1NS2A 2010 Dengue virus 2 isolate NS2BNS3NS4A2KNS 14905/BR-PE/10, 4BNS5UTR3 complete genome AGN94882 3391 2 Brazil UTR5CMENS1NS2A 2010 Dengue virus 2 isolate NS2BNS3NS4A2KNS 19190/BR-PE/10, 4BNS5UTR3 complete genome AGN94884 3391 2 Brazil UTR5CMENS1NS2A 1995 Dengue virus 2 isolate NS2BNS3NS4A2KNS 3275/BR-PE/95, 4BNS5UTR3 complete genome AGN94885 3391 2 Brazil UTR5CMENS1NS2A 1995 Dengue virus 2 isolate NS2BNS3NS4A2KNS 3311/BR-PE/95, 4BNS5UTR3 complete genome AGN94886 3391 2 Brazil UTR5CMENS1NS2A 1995 Dengue virus 2 isolate NS2BNS3NS4A2KNS 3337/BR-PE/95, 4BNS5UTR3 complete genome AGN94887 3391 2 Brazil UTR5CMENS1NS2A 1997 Dengue virus 2 isolate NS2BNS3NS4A2KNS 37473/BR-PE/97, 4BNS5UTR3 complete genome AGN94888 3391 2 Brazil UTR5CMENS1NS2A 1998 Dengue virus 2 isolate NS2BNS3NS4A2KNS 47913/BR-PE/98, 4BNS5UTR3 complete genome AGN94889 3391 2 Brazil UTR5CMENS1NS2A 1998 Dengue virus 2 isolate NS2BNS3NS4A2KNS 51347/BR-PE/98, 4BNS5UTR3 complete genome AGN94890 3391 2 Brazil UTR5CMENS1NS2A 1999 Dengue virus 2 isolate NS2BNS3NS4A2KNS 57135/BR-PE/99, 4BNS5UTR3 complete genome AGN94891 3391 2 Brazil UTR5CMENS1NS2A 2000 Dengue virus 2 isolate NS2BNS3NS4A2KNS 72144/BR-PE/00, 4BNS5UTR3 complete genome AGN94892 3391 2 Brazil UTR5CMENS1NS2A 2002 Dengue virus 2 isolate NS2BNS3NS4A2KNS 87086/BR-PE/02, 4BNS5UTR3 complete genome AGN94883 3391 2 Brazil UTR5CMENS1NS2A 2010 Dengue virus 2 isolate NS2BNS3NS4A2KNS 9479/BR-PE/10, 4BNS5UTR3 complete genome AGK36299 3391 2 Brazil CMENS1NS2ANS2B Mar. 30, 2010 Dengue virus 2 isolate NS3NS4A2KNS4BNS ACS380, complete 5UTR3 genome AGK36289 3391 2 Brazil UTR5CMENS1NS2A Mar. 1, 2010 Dengue virus 2 isolate NS2BNS3NS4A2KNS ACS46, complete 4BNS5UTR3 genome AGK36290 3391 2 Brazil UTR5CMENS1NS2A Mar. 1, 2010 Dengue virus 2 isolate NS2BNS3NS4A2KNS ACS46_II, complete 4BNS5UTR3 genome AGK36291 3391 2 Brazil UTR5CMENS1NS2A Apr. 12, 2010 Dengue virus 2 isolate NS2BNS3NS4A2KNS ACS538, complete 4BNS5UTR3 genome AGK36292 3391 2 Brazil UTR5CMENS1NS2A May 4, 2010 Dengue virus 2 isolate NS2BNS3NS4A2KNS ACS542, complete 4BNS5UTR3 genome AGK36294 3391 2 Brazil UTR5CMENS1NS2A May 4, 2010 Dengue virus 2 isolate NS2BNS3NS4A2KNS ACS721, complete 4BNS5UTR3 genome ACO06152 3391 2 Brazil UTR5CMENS1NS2A 2000 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/BR/BID- 4BNS5UTR3 V2376/2000, complete genome AET43250 3391 2 Brazil CMENS1NS2ANS2B 2000 Dengue virus 2 isolate NS3NS4A2KNS4BNS DENV-2/BR/BID- 5UTR3 V2377/2000, complete genome ACO06154 3391 2 Brazil UTR5CMENS1NS2A 2001 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/BR/BID- 4BNS5UTR3 V2379/2001, complete genome ACO06156 3391 2 Brazil UTR5CMENS1NS2A 2002 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/BR/BID- 4BNS5UTR3 V2382/2002, complete genome ACW82928 3391 2 Brazil UTR5CMENS1NS2A 2003 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/BR/BID- 4BNS5UTR3 V2385/2003, complete genome ACO06158 3391 2 Brazil UTR5CMENS1NS2A 2003 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/BR/BID- 4BNS5UTR3 V2386/2003, complete genome ACO06162 3391 2 Brazil UTR5CMENS1NS2A 2004 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/BR/BID- 4BNS5UTR3 V2390/2004, complete genome ACO06165 3391 2 Brazil UTR5CMENS1NS2A 2005 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/BR/BID- 4BNS5UTR3 V2393/2005, complete genome ACO06168 3391 2 Brazil UTR5CMENS1NS2A 2006 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/BR/BID- 4BNS5UTR3 V2396/2006, complete genome ACO06171 3391 2 Brazil UTR5CMENS1NS2A 2007 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/BR/BID- 4BNS5UTR3 V2399/2007, complete genome ACS32031 3391 2 Brazil UTR5CMENS1NS2A 2008 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/BR/BID- 4BNS5UTR3 V2402/2008, complete genome ACW82873 3391 2 Brazil UTR5CMENS1NS2A 2008 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/BR/BID- 4BNS5UTR3 V3481/2008, complete genome ACW82874 3391 2 Brazil UTR5CMENS1NS2A 2008 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/BR/BID- 4BNS5UTR3 V3483/2008, complete genome ACW82875 3391 2 Brazil UTR5CMENS1NS2A 2008 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/BR/BID- 4BNS5UTR3 V3486/2008, complete genome ACY70763 3391 2 Brazil UTR5CMENS1NS2A 2008 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/BR/BID- 4BNS5 V3495/2008, complete genome ADI80655 3391 2 Brazil UTR5CMENS1NS2A 2008 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/BR/BID- 4BNS5 V3637/2008, complete genome ACY70778 3391 2 Brazil UTR5CMENS1NS2A 2008 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/BR/BID- 4BNS5UTR3 V3638/2008, complete genome ACY70779 3391 2 Brazil UTR5CMENS1NS2A 2008 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/BR/BID- 4BNS5 V3640/2008, complete genome ACY70780 3391 2 Brazil UTR5CMENS1NS2A 2008 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/BR/BID- 4BNS5 V3644/2008, complete genome ACY70781 3391 2 Brazil UTR5CMENS1NS2A 2008 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/BR/BID- 4BNS5UTR3 V3645/2008, complete genome ACY70782 3391 2 Brazil UTR5CMENS1NS2A 2008 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/BR/BID- 4BNS5UTR3 V3648/2008, complete genome ACY70783 3391 2 Brazil UTR5CMENS1NS2A 2008 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/BR/BID- 4BNS5UTR3 V3650/2008, complete genome ACY70784 3391 2 Brazil UTR5CMENS1NS2A 2008 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/BR/BID- 4BNS5UTR3 V3653/2008, complete genome AGK36297 3391 2 Brazil UTR5CMENS1NS2A Apr. 15, 2010 Dengue virus 2 isolate NS2BNS3NS4A2KNS DGV106, complete 4BNS5UTR3 genome AGK36295 3391 2 Brazil UTR5CMENS1NS2A Feb. 24, 2010 Dengue virus 2 isolate NS2BNS3NS4A2KNS DGV34, complete 4BNS5UTR3 genome AGK36293 3391 2 Brazil UTR5CMENS1NS2A Feb. 24, 2010 Dengue virus 2 isolate NS2BNS3NS4A2KNS DGV37, complete 4BNS5UTR3 genome AGK36298 3391 2 Brazil UTR5CMENS1NS2A Mar. 9, 2010 Dengue virus 2 isolate NS2BNS3NS4A2KNS DGV69, complete 4BNS5UTR3 genome AGK36296 3391 2 Brazil UTR5CMENS1NS2A Mar. 24, 2010 Dengue virus 2 isolate NS2BNS3NS4A2KNS DGV91, complete 4BNS5UTR3 genome AFV95788 3391 2 Brazil CMENS1NS2ANS2B 2008 Dengue virus 2 strain NS3NS4A2KNS4BNS5 BR0337/2008/RJ/2008 polyprotein gene, partial cds AFV95787 3391 2 Brazil CMENS1NS2ANS2B 2008 Dengue virus 2 strain NS3NS4A2KNS4BNS5 BR0450/2008/RJ/2008 polyprotein gene, partial cds ADV39968 3391 2 Brazil CMENS1NS2ANS2B 2008 Dengue virus 2 strain NS3NS4A2KNS4BNS5 BR0690/RJ/2008 polyprotein gene, complete cds ADV71220 3391 2 Brazil CMENS1NS2ANS2B 1990 Dengue virus 2 strain NS3NS4A2KNS4BNS5 BR39145/RJ/90 polyprotein gene, partial cds ADV71215 3391 2 Brazil CMENS1NS2ANS2B 1990 Dengue virus 2 strain NS3NS4A2KNS4BNS5 BR41768/RJ/90 polyprotein gene, partial cds ADV71216 3391 2 Brazil CMENS1NS2ANS2B 1991 Dengue virus 2 strain NS3NS4A2KNS4BNS5 BR42727/RJ/91 polyprotein gene, partial cds ADV71217 3391 2 Brazil CMENS1NS2ANS2B 1994 Dengue virus 2 strain NS3NS4A2KNS4BNS5 BR48622/CE/94 polyprotein gene, partial cds ADV71218 3391 2 Brazil CMENS1NS2ANS2B 1998 Dengue virus 2 strain NS3NS4A2KNS4BNS5 BR61310/RJ/98 polyprotein gene, partial cds ADV71219 3391 2 Brazil CMENS1NS2ANS2B 1999 Dengue virus 2 strain NS3NS4A2KNS4BNS5 BR64905/RJ/99 polyprotein gene, partial cds AFH53774 3390 2 Brazil UTR5CMENS1NS2A Dengue virus 2 strain NS2BNS3NS4A2KNS JHA1, partial genome 4BNS5 AGN94893 3390 3 Brazil UTR5CMENS1NS2A 2003 Dengue virus 3 isolate NS2BNS3NS4A2KNS 101905/BR-PE/03, 4BNS5UTR3 complete genome AGN94902 3390 3 Brazil UTR5CMENS1NS2A 2004 Dengue virus 3 isolate NS2BNS3NS4A2KNS 129/BR-PE/04, 4BNS5UTR3 complete genome AGN94899 3390 3 Brazil UTR5CMENS1NS2A 2004 Dengue virus 3 isolate NS2BNS3NS4A2KNS 145/BR-PE/04, 4BNS5UTR3 complete genome AGN94903 3390 3 Brazil UTR5CMENS1NS2A 2004 Dengue virus 3 isolate NS2BNS3NS4A2KNS 161/BR-PE/04, 4BNS5UTR3 complete genome AGN94896 3390 3 Brazil UTR5CMENS1NS2A 2005 Dengue virus 3 isolate NS2BNS3NS4A2KNS 206/BR-PE/05, 4BNS5UTR3 complete genome AGN94904 3390 3 Brazil UTR5CMENS1NS2A 2005 Dengue virus 3 isolate NS2BNS3NS4A2KNS 249/BR-PE/05, 4BNS5UTR3 complete genome AGN94901 3390 3 Brazil UTR5CMENS1NS2A 2005 Dengue virus 3 isolate NS2BNS3NS4A2KNS 255/BR-PE/05, 4BNS5UTR3 complete genome AGN94905 3390 3 Brazil UTR5CMENS1NS2A 2005 Dengue virus 3 isolate NS2BNS3NS4A2KNS 263/BR-PE/05, 4BNS5UTR3 complete genome AGN94898 3390 3 Brazil UTR5CMENS1NS2A 2005 Dengue virus 3 isolate NS2BNS3NS4A2KNS 277/BR-PE/05, 4BNS5UTR3 complete genome AGN94906 3390 3 Brazil UTR5CMENS1NS2A 2005 Dengue virus 3 isolate NS2BNS3NS4A2KNS 283/BR-PE/05, 4BNS5UTR3 complete genome AGN94907 3390 3 Brazil UTR5CMENS1NS2A 2005 Dengue virus 3 isolate NS2BNS3NS4A2KNS 314/BR-PE/06, 4BNS5UTR3 complete genome AGN94897 3390 3 Brazil UTR5CMENS1NS2A 2005 Dengue virus 3 isolate NS2BNS3NS4A2KNS 339/BR-PE/05, 4BNS5UTR3 complete genome AGN94908 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS 411/BR-PE/06, 4BNS5UTR3 complete genome AGN94909 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS 418/BR-PE/06, 4BNS5UTR3 complete genome AGN94910 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS 420/BR-PE/06, 4BNS5UTR3 complete genome AGN94911 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS 423/BR-PE/06, 4BNS5UTR3 complete genome AGN94912 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS 424/BR-PE/06, 4BNS5UTR3 complete genome AGN94900 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS 603/BR-PE/06, 4BNS5UTR3 complete genome AGN94895 3390 3 Brazil UTR5CMENS1NS2A 2002 Dengue virus 3 isolate NS2BNS3NS4A2KNS 81257/BR-PE/02, 4BNS5UTR3 complete genome AGN94894 3390 3 Brazil UTR5CMENS1NS2A 2002 Dengue virus 3 isolate NS2BNS3NS4A2KNS 85469/BR-PE/02, 4BNS5UTR3 complete genome AFK83756 3390 3 Brazil UTR5CMENS1NS2A 2007 Dengue virus 3 isolate NS2BNS3NS4A2KNS D3BR/ACN/2007, 4BNS5UTR3 complete genome AFK83755 3390 3 Brazil UTR5CMENS1NS2A 2009 Dengue virus 3 isolate NS2BNS3NS4A2KNS D3BR/AL95/2009, 4BNS5UTR3 complete genome AFK83754 3390 3 Brazil UTR5CMENS1NS2A 2004 Dengue virus 3 isolate NS2BNS3NS4A2KNS D3BR/BR8/04, 4BNS5UTR3 complete genome AFK83753 3390 3 Brazil UTR5CMENS1NS2A 2002 Dengue virus 3 isolate NS2BNS3NS4A2KNS D3BR/BV4/02, 4BNS5UTR3 complete genome AFK83762 3390 3 Brazil UTR5CMENS1NS2A 2002 Dengue virus 3 isolate NS2BNS3NS4A2KNS D3BR/CU6/02, 4BNS5UTR3 complete genome AFK83759 3390 3 Brazil UTR5CMENS1NS2A 2003 Dengue virus 3 isolate NS2BNS3NS4A2KNS D3BR/MR9/03, 4BNS5UTR3 complete genome AFK83761 3390 3 Brazil UTR5CMENS1NS2A 2003 Dengue virus 3 isolate NS2BNS3NS4A2KNS D3BR/PV1/03, 4BNS5UTR3 complete genome AFK83760 3390 3 Brazil UTR5CMENS1NS2A 2002 Dengue virus 3 isolate NS2BNS3NS4A2KNS D3BR/SL3/02, 4BNS5UTR3 complete genome AHG23238 3390 3 Brazil UTR5CMENS1NS2A 2002 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5 V2383/2002, complete genome ACO06159 3390 3 Brazil UTR5CMENS1NS2A 2003 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5UTR3 V2387/2003, complete genome ACO06160 3390 3 Brazil UTR5CMENS1NS2A 2003 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5UTR3 V2388/2003, complete genome ACO06163 3390 3 Brazil UTR5CMENS1NS2A 2004 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5UTR3 V2391/2004, complete genome ACO06166 3390 3 Brazil UTR5CMENS1NS2A 2005 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5UTR3 V2394/2005, complete genome ACO06169 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5UTR3 V2397/2006, complete genome ACO06172 3390 3 Brazil UTR5CMENS1NS2A 2007 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5UTR3 V2400/2007, complete genome ACO06174 3390 3 Brazil UTR5CMENS1NS2A 2008 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5UTR3 V2403/2008, complete genome ACQ44485 3390 3 Brazil UTR5CMENS1NS2A 2001 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5UTR3 V2977/2001, complete genome ACQ44486 3390 3 Brazil UTR5CMENS1NS2A 2003 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5UTR3 V2983/2003, complete genome ACY70743 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5UTR3 V3417/2006, complete genome ACY70744 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5 V3423/2006, complete genome ACY70745 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5UTR3 V3424/2006, complete genome ACY70746 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5 V3427/2006, complete genome ACY70747 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5 V3429/2006, complete genome ACY70748 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5 V3430/2006, complete genome ACW82870 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5UTR3 V3431/2006, complete genome ACY70749 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5 V3434/2006, complete genome ACY70750 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5 V3435/2006, complete genome ACY70751 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5 V3441/2006, complete genome ACY70752 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5 V3442/2006, complete genome ACW82871 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5UTR3 V3444/2006, complete genome ACY70753 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5 V3446/2006, complete genome ACY70754 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5UTR3 V3451/2006, complete genome ACY70755 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5UTR3 V3456/2006, complete genome ACY70756 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5UTR3 V3457/2006, complete genome ACY70757 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5 V3460/2006, complete genome ACW82872 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5UTR3 V3463/2006, complete genome ACY70758 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5 V3464/2006, complete genome ACY70759 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5 V3465/2006, complete genome ACY70760 3390 3 Brazil UTR5CMENS1NS2A 2007 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5 V3469/2007, complete genome ACY70761 3390 3 Brazil UTR5CMENS1NS2A 2007 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5 V3470/2007, complete genome ACY70764 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5 V3584/2006, complete genome ACY70765 3390 3 Brazil UTR5CMENS1NS2A 2007 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5 V3585/2007, complete genome ACY70766 3390 3 Brazil UTR5CMENS1NS2A 2007 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5 V3588/2007, complete genome ACY70767 3390 3 Brazil UTR5CMENS1NS2A 2007 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5 V3589/2007, complete genome ACY70768 3390 3 Brazil UTR5CMENS1NS2A 2007 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5 V3590/2007, complete genome ACY70769 3390 3 Brazil UTR5CMENS1NS2A 2007 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5 V3591/2007, complete genome ACY70770 3390 3 Brazil UTR5CMENS1NS2A 2007 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5 V3593/2007, complete genome ACY70771 3390 3 Brazil UTR5CMENS1NS2A 2007 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5UTR3 V3597/2007, complete genome ACY70772 3390 3 Brazil UTR5CMENS1NS2A 2007 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5 V3598/2007, complete genome ACY70773 3390 3 Brazil UTR5CMENS1NS2A 2007 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5 V3601/2007, complete genome ACY70774 3390 3 Brazil UTR5CMENS1NS2A 2007 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5 V3605/2007, complete genome ACY70775 3390 3 Brazil UTR5CMENS1NS2A 2007 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5 V3606/2007, complete genome ACY70776 3390 3 Brazil UTR5CMENS1NS2A 2007 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5UTR3 V3609/2007, complete genome ACY70777 3390 3 Brazil UTR5CMENS1NS2A 2007 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/BR/BID- 4BNS5UTR3 V3615/2007, complete genome AEV42062 3390 3 Brazil UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV3/BR/D3LIMHO/ 4BNS5UTR3 2006, complete genome AGH08164 3390 3 Brazil UTR5CMENS1NS2A 2002 Dengue virus 3 strain NS2BNS3NS4A2KNS 95016/BR-PE/02, 4BNS5UTR3 complete genome AEX91754 3387 4 Brazil UTR5CMENS1NS2A Sep. 8, 2010 Dengue virus 4 isolate NS2BNS3NS4A2KNS Br246RR/10, 4BNS5UTR3 complete genome AIQ84223 3387 4 Brazil UTR5CMENS1NS2A Mar. 28, 2012 Dengue virus 4 strain NS2BNS3NS4A2KNS DENV- 4BNS5UTR3 4/MT/BR12_TVP17898/ 2012 isolate serum_12, complete genome AIQ84224 3387 4 Brazil UTR5CMENS1NS2A Mar. 30, 2012 Dengue virus 4 strain NS2BNS3NS4A2KNS DENV- 4BNS5UTR3 4/MT/BR20_TVP17906/ 2012 isolate serum_20, complete genome AIQ84225 3387 4 Brazil UTR5CMENS1NS2A Mar. 30, 2012 Dengue virus 4 strain NS2BNS3NS4A2KNS DENV- 4BNS5UTR3 4/MT/BR23_TVP17909/ 2012 isolate serum_23, complete genome AIQ84226 3387 4 Brazil UTR5CMENS1NS2A Apr. 19, 2012 Dengue virus 4 strain NS2BNS3NS4A2KNS DENV- 4BNS5UTR3 4/MT/BR24_TVP17910/ 2012 isolate serum_24, complete genome AIQ84227 3387 4 Brazil UTR5CMENS1NS2A Apr. 12, 2012 Dengue virus 4 strain NS2BNS3NS4A2KNS DENV- 4BNS5UTR3 4/MT/BR27_TVP17913/ 2012 isolate serum_27, complete genome AIQ84228 3387 4 Brazil UTR5CMENS1NS2A Apr. 19, 2012 Dengue virus 4 strain NS2BNS3NS4A2KNS DENV- 4BNS5UTR3 4/MT/BR28_TVP17914/ 2012 isolate serum_28, complete genome AIQ84220 3387 4 Brazil UTR5CMENS1NS2A Apr. 23, 2012 Dengue virus 4 strain NS2BNS3NS4A2KNS DENV- 4BNS5UTR3 4/MT/BR2_TVP17888/ 2012 isolate serum_2, complete genome AIQ84245 3387 4 Brazil UTR5CMENS1NS2A Apr. 20, 2012 Dengue virus 4 strain NS2BNS3NS4A2KNS DENV- 4BNS5UTR3 4/MT/BR33_TVP17919/ 2012 isolate serum_33, complete genome AIQ84244 3387 4 Brazil UTR5CMENS1NS2A Mar. 30, 2012 Dengue virus 4 strain NS2BNS3NS4A2KNS DENV- 4BNS5UTR3 4/MT/BR35_TVP17921/ 2012 isolate serum_35, complete genome AIQ84243 3387 4 Brazil UTR5CMENS1NS2A Apr. 3, 2012 Dengue virus 4 strain NS2BNS3NS4A2KNS DENV- 4BNS5UTR3 4/MT/BR40_TVP17926/ 2012 isolate serum_40, complete genome AIQ84242 3387 4 Brazil UTR5CMENS1NS2A Apr. 5, 2012 Dengue virus 4 strain NS2BNS3NS4A2KNS DENV- 4BNS5UTR3 4/MT/BR44_TVP17930/ 2012 isolate serum_44, complete genome AIQ84241 3387 4 Brazil UTR5CMENS1NS2A Mar. 23, 2012 Dengue virus 4 strain NS2BNS3NS4A2KNS DENV- 4BNS5UTR3 4/MT/BR47_TVP17933/ 2012 isolate serum_47, complete genome AIQ84240 3387 4 Brazil UTR5CMENS1NS2A Mar. 21, 2012 Dengue virus 4 strain NS2BNS3NS4A2KNS DENV- 4BNS5UTR3 4/MT/BR48_TVP17934/ 2012 isolate serum_48, complete genome AIQ84239 3387 4 Brazil UTR5CMENS1NS2A Mar. 12, 2012 Dengue virus 4 strain NS2BNS3NS4A2KNS DENV- 4BNS5UTR3 4/MT/BR50_TVP18148/ 2012 isolate serum_50, complete genome AIQ84238 3387 4 Brazil UTR5CMENS1NS2A Mar. 20, 2012 Dengue virus 4 strain NS2BNS3NS4A2KNS DENV- 4BNS5UTR3 4/MT/BR52_TVP17938/ 2012 isolate serum_52, complete genome AIQ84237 3387 4 Brazil UTR5CMENS1NS2A Mar. 14, 2012 Dengue virus 4 strain NS2BNS3NS4A2KNS DENV- 4BNS5UTR3 4/MT/BR53_TVP17939/ 2012 isolate serum_53, complete genome AIQ84236 3387 4 Brazil UTR5CMENS1NS2A Mar. 14, 2012 Dengue virus 4 strain NS2BNS3NS4A2KNS DENV- 4BNS5UTR3 4/MT/BR55_TVP17941/ 2012 isolate serum_55, complete genome AIQ84235 3387 4 Brazil UTR5CMENS1NS2A Mar. 14, 2012 Dengue virus 4 strain NS2BNS3NS4A2KNS DENV- 4BNS5UTR3 4/MT/BR60_TVP17946/ 2012 isolate serum_60, complete genome AIQ84234 3387 4 Brazil UTR5CMENS1NS2A Apr. 19, 2012 Dengue virus 4 strain NS2BNS3NS4A2KNS DENV- 4BNS5UTR3 4/MT/BR73_TVP17951/ 2012 isolate serum_73, complete genome AIQ84233 3387 4 Brazil UTR5CMENS1NS2A Apr. 19, 2012 Dengue virus 4 strain NS2BNS3NS4A2KNS DENV- 4BNS5UTR3 4/MT/BR76_TVP17953/ 2012 isolate serum_76, complete genome AIQ84232 3387 4 Brazil UTR5CMENS1NS2A Feb. 3, 2012 Dengue virus 4 strain NS2BNS3NS4A2KNS DENV- 4BNS5UTR3 4/MT/BR84_TVP17961/ 2012 isolate serum_84, complete genome AIQ84221 3387 4 Brazil UTR5CMENS1NS2A Apr. 23, 2012 Dengue virus 4 strain NS2BNS3NS4A2KNS DENV- 4BNS5UTR3 4/MT/BR8_TVP17894/ 2012 isolate serum_8, complete genome AIQ84231 3387 4 Brazil UTR5CMENS1NS2A Feb. 3, 2012 Dengue virus 4 strain NS2BNS3NS4A2KNS DENV- 4BNS5UTR3 4/MT/BR91_TVP17968/ 2012 isolate serum_91, complete genome AIQ84230 3387 4 Brazil UTR5CMENS1NS2A Feb. 29, 2012 Dengue virus 4 strain NS2BNS3NS4A2KNS DENV- 4BNS5UTR3 4/MT/BR92_TVP17969/ 2012 isolate serum_92, complete genome AIQ84229 3387 4 Brazil UTR5CMENS1NS2A Feb. 16, 2012 Dengue virus 4 strain NS2BNS3NS4A2KNS DENV- 4BNS5UTR3 4/MT/BR94_TVP17971/ 2012 isolate serum_94, complete genome AIQ84222 3387 4 Brazil UTR5CMENS1NS2A Apr. 18, 2012 Dengue virus 4 strain NS2BNS3NS4A2KNS DENV- 4BNS5UTR3 4/MT/BR9_TVP17895/ 2012 isolate serum_9, complete genome AEW50182 3387 4 Brazil UTR5CMENS1NS2A Mar. 26, 1982 Dengue virus 4 strain NS2BNS3NS4A2KNS H402276, complete 4BNS5 genome AFX65866 3387 4 Brazil UTR5CMENS1NS2A Jul. 17, 2010 Dengue virus 4 strain NS2BNS3NS4A2KNS H772846, complete 4BNS5UTR3 genome AFX65867 3387 4 Brazil UTR5CMENS1NS2A Jul. 18, 2010 Dengue virus 4 strain NS2BNS3NS4A2KNS H772852, complete 4BNS5UTR3 genome AEW50183 3387 4 Brazil UTR5CMENS1NS2A Jul. 21, 2010 Dengue virus 4 strain NS2BNS3NS4A2KNS H772854, complete 4BNS5 genome AFX65868 3387 4 Brazil UTR5CMENS1NS2A Aug. 20, 2010 Dengue virus 4 strain NS2BNS3NS4A2KNS H773583, complete 4BNS5UTR3 genome AFX65869 3387 4 Brazil UTR5CMENS1NS2A Aug. 24, 2010 Dengue virus 4 strain NS2BNS3NS4A2KNS H774846, complete 4BNS5UTR3 genome AFX65870 3387 4 Brazil UTR5CMENS1NS2A Nov. 10, 2010 Dengue virus 4 strain NS2BNS3NS4A2KNS H775222, complete 4BNS5UTR3 genome AFX65871 3387 4 Brazil UTR5CMENS1NS2A Jan. 12, 2011 Dengue virus 4 strain NS2BNS3NS4A2KNS H778494, complete 4BNS5UTR3 genome AFX65872 3387 4 Brazil UTR5CMENS1NS2A Jan. 11, 2011 Dengue virus 4 strain NS2BNS3NS4A2KNS H778504, complete 4BNS5UTR3 genome AFX65873 3387 4 Brazil UTR5CMENS1NS2A Jan. 20, 2011 Dengue virus 4 strain NS2BNS3NS4A2KNS H778887, complete 4BNS5UTR3 genome AFX65874 3387 4 Brazil UTR5CMENS1NS2A Jan. 14, 2011 Dengue virus 4 strain NS2BNS3NS4A2KNS H779228, complete 4BNS5UTR3 genome AFX65875 3387 4 Brazil UTR5CMENS1NS2A Jan. 24, 2011 Dengue virus 4 strain NS2BNS3NS4A2KNS H779652, complete 4BNS5UTR3 genome AFX65876 3387 4 Brazil UTR5CMENS1NS2A Nov. 29, 2010 Dengue virus 4 strain NS2BNS3NS4A2KNS H780090, complete 4BNS5UTR3 genome AFX65877 3387 4 Brazil UTR5CMENS1NS2A Nov. 21, 2010 Dengue virus 4 strain NS2BNS3NS4A2KNS H780120, complete 4BNS5UTR3 genome AFX65878 3387 4 Brazil UTR5CMENS1NS2A Jan. 29, 2011 Dengue virus 4 strain NS2BNS3NS4A2KNS H780556, complete 4BNS5UTR3 genome AFX65879 3387 4 Brazil UTR5CMENS1NS2A Jan. 29, 2011 Dengue virus 4 strain NS2BNS3NS4A2KNS H780563, complete 4BNS5UTR3 genome AFX65880 3387 4 Brazil UTR5CMENS1NS2A Jan. 13, 2011 Dengue virus 4 strain NS2BNS3NS4A2KNS H780571, complete 4BNS5UTR3 genome AFX65881 3387 4 Brazil UTR5CMENS1NS2A Mar. 18, 2011 Dengue virus 4 strain NS2BNS3NS4A2KNS H781363, complete 4BNS5UTR3 genome AIK23224 3391 2 Cuba CMENS1NS2ANS2B 1981 Dengue virus 2 isolate NS3NS4A2KNS4BNS5 Cuba_A115_1981 polyprotein gene, complete cds AIK23223 3391 2 Cuba CMENS1NS2ANS2B 1981 Dengue virus 2 isolate NS3NS4A2KNS4BNS5 Cuba_A132_1981 polyprotein gene, complete cds AIK23222 3391 2 Cuba CMENS1NS2ANS2B 1981 Dengue virus 2 isolate NS3NS4A2KNS4BNS5 Cuba_A15_1981 polyprotein gene, complete cds AIK23225 3391 2 Cuba CMENS1NS2ANS2B 1981 Dengue virus 2 isolate NS3NS4A2KNS4BNS5 Cuba_A169_1981 polyprotein gene, complete cds AIK23226 3391 2 Cuba CMENS1NS2ANS2B 1981 Dengue virus 2 isolate NS3NS4A2KNS4BNS5 Cuba_A35_1981 polyprotein gene, complete cds AAW31409 3391 2 Cuba UTR5CMENS1NS2A Dengue virus type 2 NS2BNS3NS4A2KNS strain Cuba115/97, 4BNS5UTR3 complete genome AAW31407 3391 2 Cuba UTR5CMENS1NS2A Dengue virus type 2 NS2BNS3NS4A2KNS strain Cuba13/97, 4BNS5UTR3 complete genome AAW31411 3391 2 Cuba UTR5CMENS1NS2A Dengue virus type 2 NS2BNS3NS4A2KNS strain Cuba165/97, 4BNS5UTR3 complete genome AAW31412 3391 2 Cuba UTR5CMENS1NS2A 1997 Dengue virus type 2 NS2BNS3NS4A2KNS strain Cuba205/97, 4BNS5UTR3 complete genome AAW31408 3391 2 Cuba UTR5CMENS1NS2A Dengue virus type 2 NS2BNS3NS4A2KNS strain Cuba58/97, 4BNS5UTR3 complete genome AAW31410 3391 2 Cuba UTR5CMENS1NS2A 1997 Dengue virus type 2 NS2BNS3NS4A2KNS strain Cuba89/97, 4BNS5UTR3 complete genome AFJ91714 3392 1 USA UTR5CMENS1NS2A 2010/10 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/BOL- 4BNS5UTR3 KW010, complete genome ACA48834 3392 1 USA UTR5CMENS1NS2A 1998 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V1162/1998, complete genome ACJ04186 3392 1 USA UTR5CMENS1NS2A 1995 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V1734/1995, complete genome ACJ04190 3392 1 USA UTR5CMENS1NS2A 1998 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V1738/1998, complete genome ACH99678 3392 1 USA UTR5CMENS1NS2A 1998 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V1739/1998, complete genome ACH99679 3392 1 USA UTR5CMENS1NS2A 1998 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V1740/1998, complete genome ACJ04191 3392 1 USA UTR5CMENS1NS2A 1998 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V1741/1998, complete genome ACJ04192 3392 1 USA UTR5CMENS1NS2A 1998 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V1742/1998, complete genome ACH99680 3392 1 USA UTR5CMENS1NS2A 1995 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V1743/1995, complete genome ACH99681 3392 1 USA UTR5CMENS1NS2A 1995 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V1744/1995, complete genome ACJ04215 3392 1 USA UTR5CMENS1NS2A 1998 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V2093/1998, complete genome ACJ04216 3392 1 USA UTR5CMENS1NS2A 1995 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V2094/1995, complete genome ACJ04217 3392 1 USA UTR5CMENS1NS2A 1994 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V2095/1994, complete genome ACL99012 3392 1 USA UTR5CMENS1NS2A 1993 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V2096/1993, complete genome ACL99013 3392 1 USA UTR5CMENS1NS2A 1986 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V2097/1986, complete genome ACJ04221 3392 1 USA UTR5CMENS1NS2A 1994 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V2127/1994, complete genome ACJ04222 3392 1 USA UTR5CMENS1NS2A 1995 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V2128/1995, complete genome ACJ04223 3392 1 USA UTR5CMENS1NS2A 1995 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V2129/1995, complete genome ACL99002 3392 1 USA UTR5CMENS1NS2A 1995 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V2130/1995, complete genome ACJ04224 3392 1 USA UTR5CMENS1NS2A 1996 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V2131/1996, complete genome ACJ04225 3392 1 USA UTR5CMENS1NS2A 1993 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V2132/1993, complete genome ACJ04226 3392 1 USA UTR5CMENS1NS2A 1993 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V2133/1993, complete genome ACJ04227 3392 1 USA UTR5CMENS1NS2A 1993 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V2134/1993, complete genome ACL99003 3392 1 USA UTR5CMENS1NS2A 1992 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V2135/1992, complete genome ACJ04228 3392 1 USA UTR5CMENS1NS2A 1992 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V2136/1992, complete genome ACJ04229 3392 1 USA UTR5CMENS1NS2A 1992 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V2137/1992, complete genome ACK28188 3392 1 USA UTR5CMENS1NS2A 1996 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V2138/1996, complete genome ACJ04230 3392 1 USA UTR5CMENS1NS2A 1996 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V2139/1996, complete genome ACJ04231 3392 1 USA UTR5CMENS1NS2A 1996 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V2140/1996, complete genome ACK28189 3392 1 USA UTR5CMENS1NS2A 1987 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V2142/1987, complete genome ACJ04232 3392 1 USA UTR5CMENS1NS2A 1987 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V2143/1987, complete genome ACA48858 3392 1 USA UTR5CMENS1NS2A 2006 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V852/2006, complete genome ACA48859 3392 1 USA UTR5CMENS1NS2A 1998 Dengue virus 1 isolate NS2BNS3NS4A2KNS DENV-1/US/BID- 4BNS5UTR3 V853/1998, complete genome ACF49259 3392 1 USA UTR5CMENS1NS2A 1944 Dengue virus 1 isolate NS2BNS3NS4A2KNS US/Hawaii/1944, 4BNS5UTR3 complete genome AIU47321 3392 1 USA UTR5CMENS1NS2A 1944 Dengue virus 1 strain NS2BNS3NS4A2KNS Hawaii, complete 4BNS5UTR3 genome ACA48811 3391 2 USA UTR5CMENS1NS2A 2006 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1031/2006, complete genome ACA48812 3391 2 USA UTR5CMENS1NS2A 1998 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1032/1998, complete genome ACA48813 3391 2 USA UTR5CMENS1NS2A 1998 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1033/1998, complete genome ACA48814 3391 2 USA UTR5CMENS1NS2A 1998 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1034/1998, complete genome ACA48815 3391 2 USA UTR5CMENS1NS2A 2006 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1035/2006, complete genome ACA48816 3391 2 USA UTR5CMENS1NS2A 2006 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1036/2006, complete genome ACA48817 3391 2 USA UTR5CMENS1NS2A 1998 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1038/1998, complete genome ACA48818 3391 2 USA UTR5CMENS1NS2A 2006 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1039/2006, complete genome ACA48819 3391 2 USA UTR5CMENS1NS2A 2006 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1040/2006, complete genome ACA48820 3391 2 USA UTR5CMENS1NS2A 2006 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1041/2006, complete genome ACA48821 3391 2 USA UTR5CMENS1NS2A 1998 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1042/1998, complete genome ACA48823 3391 2 USA UTR5CMENS1NS2A 2005 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1045/2005, complete genome ACA58330 3391 2 USA UTR5CMENS1NS2A 2004 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1046/2004, complete genome ACA48824 3391 2 USA UTR5CMENS1NS2A 1999 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1048/1999, complete genome ACA48827 3391 2 USA UTR5CMENS1NS2A 1998 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1052/1998, complete genome ACA48828 3391 2 USA UTR5CMENS1NS2A 1996 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1054/1996, complete genome ACB29511 3391 2 USA UTR5CMENS1NS2A 1996 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1055/1996, complete genome ACA58331 3391 2 USA UTR5CMENS1NS2A 1994 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1057/1994, complete genome ACA58332 3391 2 USA UTR5CMENS1NS2A 1994 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1058/1994, complete genome ACA48829 3391 2 USA UTR5CMENS1NS2A 1989 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1060/1989, complete genome ACD13309 3391 2 USA UTR5CMENS1NS2A 1989 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1061/1989, complete genome ACA48832 3391 2 USA UTR5CMENS1NS2A 1998 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1084/1998, complete genome ACA58337 3391 2 USA UTR5CMENS1NS2A 1994 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1085/1994, complete genome ACA58338 3391 2 USA UTR5CMENS1NS2A 1991 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1087/1991, complete genome ACB29512 3391 2 USA UTR5CMENS1NS2A 1986 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1163/1986, complete genome ACA48835 3391 2 USA UTR5CMENS1NS2A 1986 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1164/1986, complete genome ACA48836 3391 2 USA UTR5CMENS1NS2A 1987 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1165/1987, complete genome ACA48837 3391 2 USA UTR5CMENS1NS2A 1987 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1166/1987, complete genome ACA48838 3391 2 USA UTR5CMENS1NS2A 1987 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1167/1987, complete genome ACA48839 3391 2 USA UTR5CMENS1NS2A 1987 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1168/1987, complete genome ACA48840 3391 2 USA UTR5CMENS1NS2A 1987 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1169/1987, complete genome ACA48841 3391 2 USA UTR5CMENS1NS2A 1987 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1170/1987, complete genome ACA48842 3391 2 USA UTR5CMENS1NS2A 1987 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1171/1987, complete genome ACA48843 3391 2 USA UTR5CMENS1NS2A 1987 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1172/1987, complete genome ACA48844 3391 2 USA UTR5CMENS1NS2A 1987 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1174/1987, complete genome ACA48845 3391 2 USA UTR5CMENS1NS2A 1988 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1175/1988, complete genome ACA48846 3391 2 USA UTR5CMENS1NS2A 1988 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1176/1988, complete genome ACA48847 3391 2 USA UTR5CMENS1NS2A 1989 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1177/1989, complete genome ACA48848 3391 2 USA UTR5CMENS1NS2A 1989 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1178/1989, complete genome ACA48849 3391 2 USA UTR5CMENS1NS2A 1989 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1179/1989, complete genome ACA48850 3391 2 USA UTR5CMENS1NS2A 1989 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1180/1989, complete genome ACA48851 3391 2 USA UTR5CMENS1NS2A 1989 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1181/1989, complete genome ACA48852 3391 2 USA UTR5CMENS1NS2A 1989 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1182/1989, complete genome ACA48853 3391 2 USA UTR5CMENS1NS2A 1990 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1183/1990, complete genome ACA48854 3391 2 USA UTR5CMENS1NS2A 1990 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1188/1990, complete genome ACA48855 3391 2 USA UTR5CMENS1NS2A 1990 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1189/1990, complete genome ACB29513 3391 2 USA UTR5CMENS1NS2A 1993 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1356/1993, complete genome ACA48856 3391 2 USA UTR5CMENS1NS2A 1993 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1360/1993, complete genome ACB29514 3391 2 USA UTR5CMENS1NS2A 1995 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1367/1995, complete genome ACB29515 3391 2 USA UTR5CMENS1NS2A 1995 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1368/1995, complete genome ACD13310 3391 2 USA UTR5CMENS1NS2A 1995 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1372/1995, complete genome ACB29516 3391 2 USA UTR5CMENS1NS2A 1995 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1373/1995, complete genome ACB29517 3391 2 USA UTR5CMENS1NS2A 1996 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1376/1996, complete genome ACB87126 3391 2 USA UTR5CMENS1NS2A 1996 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1378/1996, complete genome ACB29518 3391 2 USA UTR5CMENS1NS2A 1996 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1383/1996, complete genome ACB87127 3391 2 USA UTR5CMENS1NS2A 1996 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1385/1996, complete genome ACD13396 3391 2 USA UTR5CMENS1NS2A 1998 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1387/1998, complete genome ACD13311 3391 2 USA UTR5CMENS1NS2A 1998 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1388/1998, complete genome ACB29519 3391 2 USA UTR5CMENS1NS2A 1998 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1392/1998, complete genome ACB29520 3391 2 USA UTR5CMENS1NS2A 1998 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1393/1998, complete genome ACB87128 3391 2 USA UTR5CMENS1NS2A 1998 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1394/1998, complete genome ACB29521 3391 2 USA UTR5CMENS1NS2A 1997 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1395/1997, complete genome ACB29522 3391 2 USA UTR5CMENS1NS2A 1997 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1396/1997, complete genome ACB29523 3391 2 USA UTR5CMENS1NS2A 1997 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1397/1997, complete genome ACB29524 3391 2 USA UTR5CMENS1NS2A 1997 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1398/1997, complete genome ACB29525 3391 2 USA UTR5CMENS1NS2A 1997 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1399/1997, complete genome ACB29526 3391 2 USA UTR5CMENS1NS2A 1997 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1401/1997, complete genome ACB29527 3391 2 USA UTR5CMENS1NS2A 1997 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1404/1997, complete genome ACB29528 3391 2 USA UTR5CMENS1NS2A 1997 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1409/1997, complete genome ACB87129 3391 2 USA UTR5CMENS1NS2A 2007 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1410/2007, complete genome ACB87130 3391 2 USA UTR5CMENS1NS2A 2007 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1411/2007, complete genome ACB87131 3391 2 USA UTR5CMENS1NS2A 2007 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1412/2007, complete genome ACB87132 3391 2 USA UTR5CMENS1NS2A 2007 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1413/2007, complete genome ACD13348 3391 2 USA UTR5CMENS1NS2A 1996 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1424/1996, complete genome ACD13349 3391 2 USA UTR5CMENS1NS2A 1999 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1425/1999, complete genome ACD13350 3391 2 USA UTR5CMENS1NS2A 1999 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1426/1999, complete genome ACD13351 3391 2 USA UTR5CMENS1NS2A 1999 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1427/1999, complete genome ACD13352 3391 2 USA UTR5CMENS1NS2A 1999 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1428/1999, complete genome ACD13353 3391 2 USA UTR5CMENS1NS2A 2004 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1431/2004, complete genome ACD13354 3391 2 USA UTR5CMENS1NS2A 2004 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1432/2004, complete genome ACD13397 3391 2 USA UTR5CMENS1NS2A 2004 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1434/2004, complete genome ACD13398 3391 2 USA UTR5CMENS1NS2A 2004 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1435/2004, complete genome ACD13399 3391 2 USA UTR5CMENS1NS2A 2004 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1436/2004, complete genome ACD13400 3391 2 USA UTR5CMENS1NS2A 2005 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1439/2005, complete genome ACE63530 3391 2 USA UTR5CMENS1NS2A 2005 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1440/2005, complete genome ACD13401 3391 2 USA UTR5CMENS1NS2A 2005 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1441/2005, complete genome ACE63543 3391 2 USA UTR5CMENS1NS2A 2005 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1442/2005, complete genome ACD13406 3391 2 USA UTR5CMENS1NS2A 2000 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1461/2000, complete genome ACD13407 3391 2 USA UTR5CMENS1NS2A 2000 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1462/2000, complete genome ACD13408 3391 2 USA UTR5CMENS1NS2A 2000 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1463/2000, complete genome ACD13409 3391 2 USA UTR5CMENS1NS2A 2000 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1464/2000, complete genome ACD13411 3391 2 USA UTR5CMENS1NS2A 2001 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1467/2001, complete genome ACD13412 3391 2 USA UTR5CMENS1NS2A 2001 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1468/2001, complete genome ACD13413 3391 2 USA UTR5CMENS1NS2A 2001 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1469/2001, complete genome ACD13414 3391 2 USA UTR5CMENS1NS2A 2001 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1470/2001, complete genome ACD13415 3391 2 USA UTR5CMENS1NS2A 2001 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1471/2001, complete genome ACD13416 3391 2 USA UTR5CMENS1NS2A 2001 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1472/2001, complete genome ACD13395 3391 2 USA UTR5CMENS1NS2A 2003 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1482/2003, complete genome ACD13419 3391 2 USA UTR5CMENS1NS2A 2003 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1483/2003, complete genome ACD13420 3391 2 USA UTR5CMENS1NS2A 2003 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1484/2003, complete genome ACD13421 3391 2 USA UTR5CMENS1NS2A 2003 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1486/2003, complete genome ACD13422 3391 2 USA UTR5CMENS1NS2A 2003 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1487/2003, complete genome ACD13424 3391 2 USA UTR5CMENS1NS2A 2003 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1492/2003, complete genome ACD13425 3391 2 USA UTR5CMENS1NS2A 2003 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1493/2003, complete genome ACD13426 3391 2 USA UTR5CMENS1NS2A 2004 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1494/2004, complete genome ACD13427 3391 2 USA UTR5CMENS1NS2A 2004 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1495/2004, complete genome ACD13428 3391 2 USA UTR5CMENS1NS2A 2004 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1496/2004, complete genome ACD13429 3391 2 USA UTR5CMENS1NS2A 2005 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V1497/2005, complete genome AEH59341 3390 2 USA CMENS1NS2ANS2B 2009 Dengue virus 2 isolate NS3NS4A2KNS4BNS5 DENV-2/US/BID- V4824/2009, complete genome AEH59346 3391 2 USA UTR5CMENS1NS2A 2006 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5 V5411/2006, complete genome AEH59345 3391 2 USA UTR5CMENS1NS2A 2007 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5 V5412/2007, complete genome ACA58343 3391 2 USA UTR5CMENS1NS2A 2006 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V585/2006, complete genome ACA48986 3391 2 USA UTR5CMENS1NS2A 2006 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V587/2006, complete genome ACA48987 3391 2 USA UTR5CMENS1NS2A 2006 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V588/2006, complete genome ACA48988 3391 2 USA UTR5CMENS1NS2A 2006 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V589/2006, complete genome ACA48989 3391 2 USA UTR5CMENS1NS2A 2002 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V591/2002, complete genome ACA48990 3391 2 USA UTR5CMENS1NS2A 2002 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V592/2002, complete genome ACA48991 3391 2 USA UTR5CMENS1NS2A 2005 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V593/2005, complete genome ACA48992 3391 2 USA UTR5CMENS1NS2A 2006 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V594/2006, complete genome ACA48993 3391 2 USA UTR5CMENS1NS2A 2006 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V595/2006, complete genome ACA48994 3391 2 USA UTR5CMENS1NS2A 1998 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V596/1998, complete genome ACA48995 3391 2 USA UTR5CMENS1NS2A 1998 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V597/1998, complete genome ACA48996 3391 2 USA UTR5CMENS1NS2A 1999 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V598/1999, complete genome ACA48997 3391 2 USA UTR5CMENS1NS2A 1999 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V599/1999, complete genome ACA48998 3391 2 USA UTR5CMENS1NS2A 2005 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V600/2005, complete genome ACA48999 3391 2 USA UTR5CMENS1NS2A 1998 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V675/1998, complete genome ACA49000 3391 2 USA UTR5CMENS1NS2A 1998 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V676/1998, complete genome ACA49001 3391 2 USA UTR5CMENS1NS2A 1998 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V677/1998, complete genome ACA49002 3391 2 USA UTR5CMENS1NS2A 1998 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V678/1998, complete genome ACA49003 3391 2 USA UTR5CMENS1NS2A 1994 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V679/1994, complete genome ACA49004 3391 2 USA UTR5CMENS1NS2A 1994 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V680/1994, complete genome ACA49005 3391 2 USA UTR5CMENS1NS2A 1998 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V681/1998, complete genome ACA49006 3391 2 USA UTR5CMENS1NS2A 1994 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V682/1994, complete genome ACA49007 3391 2 USA UTR5CMENS1NS2A 1994 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V683/1994, complete genome ACA49008 3391 2 USA UTR5CMENS1NS2A 1994 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V684/1994, complete genome ACA49009 3391 2 USA UTR5CMENS1NS2A 1988 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V685/1988, complete genome ACA49010 3391 2 USA UTR5CMENS1NS2A 1989 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V686/1989, complete genome ACA49011 3391 2 USA UTR5CMENS1NS2A 1989 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V687/1989, complete genome ACA49012 3391 2 USA UTR5CMENS1NS2A 1989 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V688/1989, complete genome ACA49013 3391 2 USA UTR5CMENS1NS2A 1989 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V689/1989, complete genome ACA49014 3391 2 USA UTR5CMENS1NS2A 1988 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V690/1988, complete genome ACA48857 3391 2 USA UTR5CMENS1NS2A 1990 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V851/1990, complete genome ACA48860 3391 2 USA UTR5CMENS1NS2A 2001 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V854/2001, complete genome ACA48861 3391 2 USA UTR5CMENS1NS2A 1992 Dengue virus 2 isolate NS2BNS3NS4A2KNS DENV-2/US/BID- 4BNS5UTR3 V855/1992, complete genome ACA48822 3390 3 USA UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1043/2006, complete genome ACA58329 3390 3 USA UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1044/2006, complete genome ACA48825 3390 3 USA UTR5CMENS1NS2A 1998 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1049/1998, complete genome ACA48826 3390 3 USA UTR5CMENS1NS2A 1998 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1050/1998, complete genome ACA48830 3390 3 USA UTR5CMENS1NS2A 1998 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1075/1998, complete genome ACA58333 3390 3 USA UTR5CMENS1NS2A 1999 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1076/1999, complete genome ACA58334 3390 3 USA UTR5CMENS1NS2A 2000 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1077/2000, complete genome ACA48831 3390 3 USA UTR5CMENS1NS2A 2003 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1078/2003, complete genome ACA58335 3390 3 USA UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1079/2006, complete genome ACA58336 3390 3 USA UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1080/2006, complete genome ACA48833 3390 3 USA UTR5CMENS1NS2A 1998 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1088/1998, complete genome ACA58339 3390 3 USA UTR5CMENS1NS2A 2003 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1089/2003, complete genome ACA58340 3390 3 USA UTR5CMENS1NS2A 1998 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1090/1998, complete genome ACA58341 3390 3 USA UTR5CMENS1NS2A 2004 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1091/2004, complete genome ACA58342 3390 3 USA UTR5CMENS1NS2A 2004 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1092/2004, complete genome ACB87133 3390 3 USA UTR5CMENS1NS2A 2007 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1415/2007, complete genome ACB87134 3390 3 USA UTR5CMENS1NS2A 2007 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1416/2007, complete genome ACB87135 3390 3 USA UTR5CMENS1NS2A 2007 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1417/2007, complete genome ACD13402 3390 3 USA UTR5CMENS1NS2A 1998 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1447/1998, complete genome ACE63531 3390 3 USA UTR5CMENS1NS2A 1998 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1448/1998, complete genome ACE63532 3390 3 USA UTR5CMENS1NS2A 1998 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1449/1998, complete genome ACH99660 3390 3 USA UTR5CMENS1NS2A 1998 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1450/1998, complete genome ACE63544 3390 3 USA UTR5CMENS1NS2A 1999 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1451/1999, complete genome ACE63545 3390 3 USA UTR5CMENS1NS2A 1999 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1452/1999, complete genome ACE63533 3390 3 USA UTR5CMENS1NS2A 1999 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1453/1999, complete genome ACE63534 3390 3 USA UTR5CMENS1NS2A 1999 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1454/1999, complete genome ACD13403 3390 3 USA UTR5CMENS1NS2A 1999 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1455/1999, complete genome ACD13405 3390 3 USA UTR5CMENS1NS2A 2000 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1460/2000, complete genome ACE63528 3390 3 USA UTR5CMENS1NS2A 2000 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1465/2000, complete genome ACD13410 3390 3 USA UTR5CMENS1NS2A 1999 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1466/1999, complete genome ACD13417 3391 3 USA UTR5CMENS1NS2A 2002 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1473/2002, complete genome ACD13418 3390 3 USA UTR5CMENS1NS2A 2002 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1475/2002, complete genome ACD13392 3390 3 USA UTR5CMENS1NS2A 2002 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1476/2002, complete genome ACH61690 3390 3 USA UTR5CMENS1NS2A 2002 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1477/2002, complete genome ACJ04182 3390 3 USA UTR5CMENS1NS2A 2002 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1478/2002, complete genome ACD13393 3390 3 USA UTR5CMENS1NS2A 2003 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1480/2003, complete genome ACD13394 3390 3 USA UTR5CMENS1NS2A 2003 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1481/2003, complete genome ACE63529 3390 3 USA UTR5CMENS1NS2A 2003 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1490/2003, complete genome ACD13423 3390 3 USA UTR5CMENS1NS2A 2003 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1491/2003, complete genome ACH99651 3390 3 USA UTR5CMENS1NS2A 2004 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1604/2004, complete genome ACO06143 3390 3 USA UTR5CMENS1NS2A 2004 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1605/2004, complete genome ACH61715 3390 3 USA UTR5CMENS1NS2A 2004 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1606/2004, complete genome ACH61716 3390 3 USA UTR5CMENS1NS2A 2004 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1607/2004, complete genome ACH61717 3390 3 USA UTR5CMENS1NS2A 2004 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1608/2004, complete genome ACH61718 3390 3 USA UTR5CMENS1NS2A 2004 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1609/2004, complete genome ACH61719 3390 3 USA UTR5CMENS1NS2A 2004 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1610/2004, complete genome ACO06144 3390 3 USA UTR5CMENS1NS2A 2004 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1611/2004, complete genome ACH61720 3390 3 USA UTR5CMENS1NS2A 2004 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1612/2004, complete genome ACH61721 3390 3 USA UTR5CMENS1NS2A 2004 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1613/2004, complete genome ACH99652 3390 3 USA UTR5CMENS1NS2A 2004 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1614/2004, complete genome ACJ04178 3390 3 USA UTR5CMENS1NS2A 2004 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1615/2004, complete genome ACH99653 3390 3 USA UTR5CMENS1NS2A 2004 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1616/2004, complete genome ACH99654 3390 3 USA UTR5CMENS1NS2A 2005 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1617/2005, complete genome ACH99655 3390 3 USA UTR5CMENS1NS2A 2005 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1618/2005, complete genome ACH99656 3390 3 USA UTR5CMENS1NS2A 2005 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1619/2005, complete genome ACH99657 3390 3 USA UTR5CMENS1NS2A 2005 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1620/2005, complete genome ACH99658 3390 3 USA UTR5CMENS1NS2A 2005 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1621/2005, complete genome ACH99665 3390 3 USA UTR5CMENS1NS2A 2005 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1622/2005, complete genome ACH99666 3390 3 USA UTR5CMENS1NS2A 2005 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1623/2005, complete genome ACH99667 3390 3 USA UTR5CMENS1NS2A 2005 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1624/2005, complete genome ACH99668 3390 3 USA UTR5CMENS1NS2A 2005 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1625/2005, complete genome ACH99669 3390 3 USA UTR5CMENS1NS2A 2005 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1626/2005, complete genome ACJ04183 3390 3 USA UTR5CMENS1NS2A 2003 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1729/2003, complete genome ACJ04184 3390 3 USA UTR5CMENS1NS2A 2003 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1730/2003, complete genome ACH99676 3390 3 USA UTR5CMENS1NS2A 2003 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1731/2003, complete genome ACJ04185 3390 3 USA UTR5CMENS1NS2A 2002 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1732/2002, complete genome ACH99677 3390 3 USA UTR5CMENS1NS2A 1999 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1733/1999, complete genome ACJ04187 3390 3 USA UTR5CMENS1NS2A 1999 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1735/1999, complete genome ACJ04188 3390 3 USA UTR5CMENS1NS2A 1999 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1736/1999, complete genome ACJ04189 3390 3 USA UTR5CMENS1NS2A 1999 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V1737/1999, complete genome ACL98985 3390 3 USA UTR5CMENS1NS2A 1999 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V2098/1999, complete genome ACL98986 3390 3 USA UTR5CMENS1NS2A 1998 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V2099/1998, complete genome ACL99014 3390 3 USA UTR5CMENS1NS2A 2000 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V2100/2000, complete genome ACL98987 3390 3 USA UTR5CMENS1NS2A 2000 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V2103/2000, complete genome ACJ04218 3390 3 USA UTR5CMENS1NS2A 2000 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V2104/2000, complete genome ACJ04219 3390 3 USA UTR5CMENS1NS2A 2000 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V2105/2000, complete genome ACL98988 3390 3 USA UTR5CMENS1NS2A 2000 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V2106/2000, complete genome ACL98989 3390 3 USA UTR5CMENS1NS2A 2000 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V2107/2000, complete genome ACL98990 3390 3 USA UTR5CMENS1NS2A 2000 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V2108/2000, complete genome ACL98991 3390 3 USA UTR5CMENS1NS2A 2000 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V2110/2000, complete genome ACL98992 3390 3 USA UTR5CMENS1NS2A 2000 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V2111/2000, complete genome ACL98993 3390 3 USA UTR5CMENS1NS2A 2000 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V2112/2000, complete genome ACL98994 3390 3 USA UTR5CMENS1NS2A 2000 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V2113/2000, complete genome ACL98995 3390 3 USA UTR5CMENS1NS2A 2001 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V2114/2001, complete genome ACL98996 3390 3 USA UTR5CMENS1NS2A 2001 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V2115/2001, complete genome ACL98997 3390 3 USA UTR5CMENS1NS2A 2001 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V2117/2001, complete genome ACL98998 3390 3 USA UTR5CMENS1NS2A 2001 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V2118/2001, complete genome ACL98999 3390 3 USA UTR5CMENS1NS2A 2002 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V2119/2002, complete genome ACJ04220 3390 3 USA UTR5CMENS1NS2A 2002 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V2120/2002, complete genome ACL99000 3390 3 USA UTR5CMENS1NS2A 2002 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V2122/2002, complete genome ACK28187 3390 3 USA UTR5CMENS1NS2A 2002 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V2123/2002, complete genome ACL99001 3390 3 USA UTR5CMENS1NS2A 2006 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V2126/2006, complete genome ACA48862 3390 3 USA UTR5CMENS1NS2A 2003 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V858/2003, complete genome ACA48863 3390 3 USA UTR5CMENS1NS2A 1998 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV-3/US/BID- 4BNS5UTR3 V859/1998, complete genome AFZ40124 3390 3 USA UTR5CMENS1NS2A 1963 Dengue virus 3 isolate NS2BNS3NS4A2KNS DENV- 4BNS5UTR3 3/USA/633798/1963, complete genome ACH61714 3387 4 USA UTR5CMENS1NS2A 1998 Dengue virus 4 isolate NS2BNS3NS4A2KNS DENV-4/US/BID- 4BNS5UTR3 V1082/1998, complete genome ACH61687 3387 4 USA UTR5CMENS1NS2A 1986 Dengue virus 4 isolate NS2BNS3NS4A2KNS DENV-4/US/BID- 4BNS5UTR3 V1083/1986, complete genome ACH61688 3387 4 USA UTR5CMENS1NS2A 1998 Dengue virus 4 isolate NS2BNS3NS4A2KNS DENV-4/US/BID- 4BNS5UTR3 V1093/1998, complete genome ACH61689 3387 4 USA UTR5CMENS1NS2A 1998 Dengue virus 4 isolate NS2BNS3NS4A2KNS DENV-4/US/BID- 4BNS5UTR3 V1094/1998, complete genome ACS32012 3387 4 USA UTR5CMENS1NS2A 1994 Dengue virus 4 isolate NS2BNS3NS4A2KNS DENV-4/US/BID- 4BNS5UTR3 V2429/1994, complete genome ACS32013 3387 4 USA UTR5CMENS1NS2A 1994 Dengue virus 4 isolate NS2BNS3NS4A2KNS DENV-4/US/BID- 4BNS5UTR3 V2430/1994, complete genome ACS32014 3387 4 USA UTR5CMENS1NS2A 1995 Dengue virus 4 isolate NS2BNS3NS4A2KNS DENV-4/US/BID- 4BNS5UTR3 V2431/1995, complete genome ACS32037 3387 4 USA UTR5CMENS1NS2A 1995 Dengue virus 4 isolate NS2BNS3NS4A2KNS DENV-4/US/BID- 4BNS5UTR3 V2432/1995, complete genome ACO06140 3387 4 USA UTR5CMENS1NS2A 1995 Dengue virus 4 isolate NS2BNS3NS4A2KNS DENV-4/US/BID- 4BNS5UTR3 V2433/1995, complete genome ACO06145 3387 4 USA UTR5CMENS1NS2A 1995 Dengue virus 4 isolate NS2BNS3NS4A2KNS DENV-4/US/BID- 4BNS5UTR3 V2434/1995, complete genome ACS32015 3387 4 USA UTR5CMENS1NS2A 1996 Dengue virus 4 isolate NS2BNS3NS4A2KNS DENV-4/US/BID- 4BNS5UTR3 V2435/1996, complete genome ACS32016 3387 4 USA UTR5CMENS1NS2A 1996 Dengue virus 4 isolate NS2BNS3NS4A2KNS DENV-4/US/BID- 4BNS5UTR3 V2436/1996, complete genome ACS32017 3387 4 USA UTR5CMENS1NS2A 1996 Dengue virus 4 isolate NS2BNS3NS4A2KNS DENV-4/US/BID- 4BNS5UTR3 V2437/1996, complete genome ACS32018 3387 4 USA UTR5CMENS1NS2A 1996 Dengue virus 4 isolate NS2BNS3NS4A2KNS DENV-4/US/BID- 4BNS5UTR3 V2438/1996, complete genome ACS32019 3387 4 USA UTR5CMENS1NS2A 1996 Dengue virus 4 isolate NS2BNS3NS4A2KNS DENV-4/US/BID- 4BNS5UTR3 V2439/1996, complete genome ACO06146 3387 4 USA UTR5CMENS1NS2A 1996 Dengue virus 4 isolate NS2BNS3NS4A2KNS DENV-4/US/BID- 4BNS5UTR3 V2440/1996, complete genome ACQ44402 3387 4 USA UTR5CMENS1NS2A 1998 Dengue virus 4 isolate NS2BNS3NS4A2KNS DENV-4/US/BID- 4BNS5UTR3 V2441/1998, complete genome ACQ44403 3387 4 USA UTR5CMENS1NS2A 1998 Dengue virus 4 isolate NS2BNS3NS4A2KNS DENV-4/US/BID- 4BNS5UTR3 V2442/1998, complete genome ACO06147 3387 4 USA UTR5CMENS1NS2A 1998 Dengue virus 4 isolate NS2BNS3NS4A2KNS DENV-4/US/BID- 4BNS5UTR3 V2443/1998, complete genome ACQ44404 3387 4 USA UTR5CMENS1NS2A 1998 Dengue virus 4 isolate NS2BNS3NS4A2KNS DENV-4/US/BID- 4BNS5UTR3 V2444/1998, complete genome ACQ44405 3387 4 USA UTR5CMENS1NS2A 1998 Dengue virus 4 isolate NS2BNS3NS4A2KNS DENV-4/US/BID- 4BNS5UTR3 V2445/1998, complete genome ACQ44406 3387 4 USA UTR5CMENS1NS2A 1999 Dengue virus 4 isolate NS2BNS3NS4A2KNS DENV-4/US/BID- 4BNS5UTR3 V2446/1999, complete genome ACQ44407 3387 4 USA UTR5CMENS1NS2A 1999 Dengue virus 4 isolate NS2BNS3NS4A2KNS DENV-4/US/BID- 4BNS5UTR3 V2447/1999, complete genome ACQ44408 3387 4 USA UTR5CMENS1NS2A 1999 Dengue virus 4 isolate NS2BNS3NS4A2KNS DENV-4/US/BID- 4BNS5UTR3 V2448/1999, complete genome ACJ04171 3387 4 USA UTR5CMENS1NS2A 1994 Dengue virus 4 isolate NS2BNS3NS4A2KNS DENV-4/US/BID- 4BNS5UTR3 V860/1994, complete genome

TABLE 43 mRNA constructs that encode one or more OVA epitopes #of Antigen Peptides/ Presentation SEQ ID Construct Repeats Linker Enhancer Sequence Amino acid Sequence NO:  1 8 OVA G/S — MLESIINFEKLTEGGGGSG 345 (8mer) GGGSLESIINFEKLTEGGG Repeats GSGGGGSLESIINFEKLTE (Flanking GGGGSGGGGSLESIINFEK AA) LTEGGGGSGGGGSLESIIN FEKLTEGGGGSGGGGSLE SIINFEKLTEGGGGSGGG GSLESIINFEKLTEGGGGS GGGGSLESIINFEKLTE  2 8 OVA Cathepsin — MLESIINFEKLTEGFLGLE 346 (8mer) B SIINFEKLTEGFLGLESIIN Repeats Cleavage FEKLTEGFLGLESIINFEK (Flanking Site LTEGFLGLESIINFEKLTE AA) (GFLG) GFLGLESIINFEKLTEGFL GLESIINFEKLTEGFLGLE SIINFEKLTE  3 8 OVA — Human MHCI MRVTAPRTVLLLLSAALA 347 (8mer) Secretion LTETWALESIINFEKLTEL Repeats Peptide/ ESIINFEKLTELESIINFEKL (Flanking Cytoplasmic TELESIINFEKLTELESIINF AA) Domain EKLTELESIINFEKLTELES IINFEKLTELESIINFEKLT EGSIVGIVAGLAVLAVVV IGAVVATVMCRRKSSGG KGGSYSQAASSDSAQGS DVSLTA  4 8 OVA Cathepsin Human MHCI MRVTAPRTVLLLLSAALA 348 (8mer) B Secretion LTETWALESIINFEKLTEG Repeats Cleavage Peptide/ FLGLESIINFEKLTEGFLG (Flanking Site Cytoplasmic LESIINFEKLTEGFLGLESII AA) (GFLG) Domain NFEKLTEGFLGLESIINFE KLTEGFLGLESIINFEKLT EGFLGLESIINFEKLTEGF LGLESIINFEKLTEGSIVGI VAGLAVLAVVVIGAVVA TVMCRRKSSGGKGGSYS QAASSDSAQGSDVSLTA  5 Single OVA — KDEL MSIINFEKLKDEL 349  6 Single OVA — Human MHCI MRVTAPRTVLLLLSAALA 350 (Flanking Secretion LTETWALESIINFEKLTEG AA) Peptide/ SIVGIVAGLAVLAVVVIG Cytoplasmic AVVATVMCRRKSSGGKG Domain GSYSQAASSDSAQGSDVS LTA  7 8 OVA Cathepsin Murine Ig Kappa METDTLLLWVLLLWVPG 351 (8mer) B Signal Peptide STGDSIINFEKLGFLGSIIN Repeats Cleavage (Igκ) PEKLGFLGSIINFEKLGFL Site GSIINFEKLGFLGSIINFEK (GFLG) LGFLGSIINFEKLGFLGSII NFEKLGFLGSIINFEKL  8 8 OVA G/S Human MHCI MRVTAPRTVLLLLSAALA 352 (8mer) Secretion LTETWALESIINFEKLTEG Repeats Peptide/ GGGSGGGGSLESIINFEKL (Flanking Cytoplasmic TEGGGGSGGGGSLESIINF AA) Domain EKLTEGGGGSGGGGSLES IINFEKLTEGGGGSGGGG SLESIINFEKLTEGGGGSG GGGSLESIINFEKLTEGGG GSGGGGSLESIINFEKLTE GGGGSGGGGSLESIINFEK LTEGSIVGIVAGLAVLAV VVIGAVVATVMCRRKSS GGKGGSYSQAASSDSAQ GSDVSLTA  9 8 OVA — — MLESIINFEKLTELESIINF 353 (8mer) EKLTELESIINFEKLTELES Repeats IINFEKLTELESIINFEKLT (Flanking ELESIINFEKLTELESIINFE AA) KLTELESIINFEKLTE 10 Single OVA — — MSIINFEKL 354 11 8 OVA Cathepsin Murine Ig Kappa METDTLLLWVLLLWVPG 355 (8mer) B Signal Peptide STGDHPFTEDDAVDPNDS Repeats Cleavage (Igκ) and PEST DIDPESRSIINFEKLGFLGS Site IINFEKLGFLGSIINFEKLG (GFLG) FLGSIINFEKLGFLGSIINF EKLGFLGSIINFEKLGFLG SIINFEKLGFLGSIINFEKL 12 8 OVA Cathepsin Marine MHC Class MSIINFEKLGFLGSIINFEK 356 (8mer) B I Cytoplasmic  LGFLGSIINFEKLGFLGSII Repeats Cleavage Domain (MITD) NFEKLGFLGSIINFEKLGF Site LGSIINFEKLGFLGSIINFE (GFLG) KLGFLGSIINFEKLPPPSTV SNMIIIEVLIVLGAVINIGAM VAFVLKSKRKIGGKGGVYA LAGGSNSIHGSALFLEAFK A

TABLE 44 DENV mRNA vaccine constructs tested for antibody binding or in challenge studies Construct mRNA Name 13 DEN3_prME_PaH881/88_AF349753.1 14 DEN1_prME_West_Pac_AY145121.1 15 DEN1_prME_PUO-359_AAN32784.1 16 DEN4_prME_DHF_Patient_JN638571.1 17 DEN4_prME_DENV4/CN/GZ29/2010_KP723482.1 18 DEN4_prME_rDEN4_AF326825.1 19 DEN3_prME_L11439.1 20 DEN3_prME_D3/Hu/TL129NIID/2005_AB214882 21 DENV2_prME_Peru_IQT2913_1996 22 DENV2_prME_Thailand-168_1979 23 DENV2_prME_Thailand_PUO-218_1980 (Sanofi strain) 24 DEN2_D2Y98P_PRME80_Hs3_LSP 25 Non-H2Kb multitope 26 H2Kb multitope

TABLE 45 DENV prME Challenge Study Design in Cynomolgus (cyno) Monkey Group Vaccine n = 3 Vaccine Schedule Dosage/Route Challenge 1 Dengue 1 Day 0, 21, 42 IM, LNP Challenge with Dengue prME 250 μg 1/03135 s.c (5log PFU) 2 (Construct 15) IM, LNP 5 μg 3 Dengue 2 Day 0, 21, 42 IM, LNP Challenge with Dengue prME 250 μg 2/99345 s.c (5log PFU) 4 (Construct 21) IM, LNP 5 μg 5 Dengue 3 Day 0, 21, 42 IM, LNP Challenge with Dengue prME 250 μg 3/16562 s.c (5log PFU) 6 (Construct 19) IM, LNP 5 μg 7 Dengue 4 Day 0, 21, 42 IM, LNP Challenge with Dengue prME 250 μg 4/1036 s.c (5log PFU) 8 (Construct 17) IM, LNP 5 μg 9 prME Combo Day 0, 21, 42 IM, LNP Challenge with Dengue (Post- 1000 μg Total (250 μg 1/03135 s.c (5log PFU) Formulation of each) 10 Mix) IM, LNP (Constructs 15, 20 μg Total (5 μg 17, 19, and 21) of each) 11 prME Combo Day 0, 21, 42 IM, LNP Challenge with Dengue (Post- 1000 μg Total (250 μg 2/99345 s.c (5log PFU) Formulation of each) 12 Mix) IM, LNP (Constructs 15, 20 μg Total (5 μg 17, 19, and 21) of each) 13 prME Combo Day 0, 21, 42 IM, LNP Challenge with Dengue (Post- 1000 μg Total (250 μg 3/16562 s.c (5log PFU) Formulation of each) 14 Mix) IM, LNP (Constructs 15, 20 μg Total (5 μg 17, 19, and 21) of each) 15 prME Combo Day 0, 21, 42 IM, LNP Challenge with Dengue (Post- 1000 μg Total (250 μg 4/1036 s.c (5log PFU) Formulation of each) 16 Mix) IM, LNP (Constructs 15, 20 μg Total (5 μg 17, 19, and 21) of each) 17 prME Combo Day 0, 21, 42 IM, LNP Challenge with Dengue (Post- 1000 μg Total (250 μg 2/99345 s.c (5log PFU) Formulation of each) Mix) (Constructs 15, 17, 19, and 22) 18 H10N8-FLAG Day 0, 21, 42 IM, LNP Challenge with Dengue 250 μg 2/99345 s.c (5log PFU) 19 Naive — — Challenge with Dengue 1/03135 s.c (5log PFU) 20 Naive — — Challenge with Dengue 2/99345 s.c (5log PFU) 21 Naive — — Challenge with Dengue 3/16562 s.c (5log PFU) 22 Naive — — Challenge with Dengue 4/1036 s.c (5log PFU) Collect serum on day 20, 41, 62, and 92 for serotype-specific neutralization assay Collect serum on day 62 (pre-challenge), 63-66, 68, 70, 72, 76, and 92 (end of In-life) to determine serum viral load

TABLE 46 DENV prME Challenge Study Design in AG129 Mice Group Vaccine n = 5 Vaccine Schedule Dosage/Route Serum/PBMCs Challenge Readout 1 Dengue 1 prME Day 0, 21 IM, LNP, 10 μg Collect serum on Challenge with Monitor 2 (Construct 15) Day 0, 21 IM, LNP, 2 μg day 20 and 41 for 1e5 PFU per weights 3 Dengue 2 prME Day 0, 21 IM, LNP, 10 μg serotype-specific mouse of and health 4 (Construct 21) Day 0, 21 IM, LNP, 2 μg neutralization assay D2Y98P SC for 14 days 5 Dengue 3 prME Day 0, 21 IM, LNP, 10 μg injection (Day p.i. 6 (Construct 19) Day 0, 21 IM, LNP, 2 μg 42) 7 Dengue 4 prME Day 0, 21 IM, LNP, 10 μg 8 (Construct 17) Day 0, 21 IM, LNP, 2 μg 9 H2Kb Multitope Day 0, 21 IM, LNP, 10 μg Collect and 10 (Construct 25) Day 0, 21 IM, LNP, 2 μg cryopreserve 11 Non-H2Kb Day 0, 21 IM, LNP, 10 μg PBMCs on day 20 12 Multitope Day 0, 21 IM, LNP, 2 μg and 41; Ship to (Construct 26) Valera 13 prME Combo + Day 0, 21 IM, LNP, 10 μg Collect serum on H2Kb Multitope Total (2 μg day 20 and 41 for (Constructs 15, of each) serotype-specific 17, 19, and 21) neutralization assay (Post7) 14 prME Combo + Day 0, 21 IM, LNP, 10 μg non-H2Kb Total (2 μg Multitope of each) (Constructs 15, 17, 19, 21, and 26) (Post7) 15 prME Combo Day 0, 21 IM, LNP, 8 μg (Constructs 15, Total (2 μg of 17, 19, and 21) each) (Post7) 16 prME Combo + Day 0, 21 IM, LNP, 10 μg H2Kb Multitope Total (2 μg (Constructs 15, of each) 17, 19, 21 and 25) (Post1) 17 prME Combo + Day 0, 21 IM, LNP, 10 μg non-H2Kb Total (2 μg Multitope of each) (Constructs 15, 17, 19, 21, and 26) (Post1) 18 prME Combo Day 0, 21 IM, LNP, 8 μg (Constructs 15, Total (2 μg of 17, 19, and 21) each) (Post1) 19 Dengue 2 prME Day 0, 21 IM, LNP, 2 μg Collect serum on (Construct 22) day 20 and 41 for 20 Naive Day 0, 21 Tris/Sucrose Dengue 2-specific neutralization assay

TABLE 47 CHIKV Polynucleotide Sequences SEQ ID Name Sequence NO: ChiK.secE1 TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGA 376 HS3UPCRfree AATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGG (CHIKV secreted AGACACCTGCACAGCTGTTGTTTCTGCTGCTGCTTTGGTTGCCCGATACC E1 antigen) ACCGGTGACTACAAAGACGACGACGATAAATACGAGCACGTGACGGTA ATACCAAACACTGTGGGGGTGCCATACAAGACCCTGGTAAATCGCCCA GGCTACTCTCCCATGGTGCTGGAGATGGAGCTCCAGTCTGTGACCTTAG AGCCAACCCTCTCACTCGACTATATCACCTGTGAATACAAAACAGTGAT CCCATCCCCCTACGTGAAATGTTGCGGAACTGCAGAGTGTAAGGATAA GAGTCTGCCCGATTACAGCTGCAAGGTGTTTACAGGCGTGTATCCATTT ATGTGGGGAGGAGCCTACTGTTTTTGCGATGCCGAAAATACTCAGCTGT CTGAAGCCCATGTGGAGAAGAGTGAAAGTTGCAAGACCGAATTTGCTA GTGCCTACAGGGCACACACCGCTTCTGCCTCCGCTAAACTCCGAGTCCT TTACCAGGGCAATAATATTACGGTCGCTGCCTACGCTAACGGGGACCAC GCTGTGACAGTCAAGGACGCCAAATTCGTAGTGGGCCCAATGAGCTCC GCCTGGACTCCCTTCGACAACAAAATCGTCGTGTATAAAGGCGACGTGT ACAATATGGACTACCCACCCTTCGGGGCTGGAAGACCGGGCCAGTTTG GAGATATCCAATCAAGGACACCCGAGTCAAAGGACGTGTACGCCAATA CGCAGCTGGTGCTGCAGAGACCCGCCGCTGGTACCGTGCATGTGCCTTA TTCCCAAGCTCCATCTGGCTTCAAGTACTGGTTGAAAGAGCGCGGTGCT TCGCTGCAGCATACAGCACCGTTCGGATGTCAGATAGCAACCAACCCTG TACGGGCTGTCAACTGTGCCGTGGGAAATATACCTATTTCCATCGACAT TCCGGACGCAGCTTTCACACGTGTCGTTGATGCCCCCTCAGTGACTGAC ATGTCATGTGAGGTGCCTGCTTGCACCCACAGCAGCGATTTTGGCGGAG TGGCCATAATCAAGTACACCGCCTCCAAAAAAGGAAAGTGTGCCGTAC ACTCTATGACCAACGCCGTCACAATCAGAGAAGCCGACGTTGAAGTAG AGGGAAATTCACAGCTGCAAATCAGCTTCAGCACCGCTCTTGCCTCTGC TGAGTTTAGGGTTCAGGTTTGCAGTACTCAGGTGCACTGTGCAGCCGCT TGCCATCCCCCCAAGGATCATATCGTGAATTATCCTGCATCCCACACCA CACTGGGAGTCCAGGATATCTCAACAACTGCAATGTCTTGGGTGCAGAA GATCACCTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCT TGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCGTGG TCTTTGAATAAAGTCTGAGTGGGCGGC Chik- TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGA 377 Strain37997-E1 AATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGT (CHIKV E1 ACGAACACGTAACAGTGATCCCGAACACGGTGGGAGTACCGTATAAGA antigen-Strain CTCTAGTCAACAGACCGGGCTACAGCCCCATGGTATTGGAGATGGAGCT 37997): TCTGTCTGTCACCTTGGAACCAACGCTATCGCTTGATTACATCACGTGC GAGTATAAAACCGTTATCCCGTCTCCGTACGTGAAATGCTGCGGTACAG CAGAGTGTAAGGACAAGAGCCTACCTGATTACAGCTGTAAGGTCTTCAC CGGCGTCTACCCATTCATGTGGGGCGGCGCCTACTGCTTCTGCGACACC GAAAATACGCAATTGAGCGAAGCACATGTGGAGAAGTCCGAATCATGC AAAACAGAATTTGCATCAGCATACAGGGCTCATACCGCATCCGCATCA GCTAAGCTCCGCGTCCTTTACCAAGGAAATAATATCACTGTGGCTGCTT ATGCAAACGGCGACCATGCCGTCACAGTTAAGGACGCTAAATTCATAG TGGGGCCAATGTCTTCAGCCTGGACACCTTTCGACAATAAAATCGTGGT GTACAAAGGCGACGTCTACAACATGGACTACCCGCCCTTCGGCGCAGG AAGACCAGGACAATTTGGCGACATCCAAAGTCGCACGCCTGAGAGCGA AGACGTCTATGCTAATACACAACTGGTACTGCAGAGACCGTCCGCGGGT ACGGTGCACGTGCCGTACTCTCAGGCACCATCTGGCTTCAAGTATTGGC TAAAAGAACGAGGGGCGTCGCTGCAGCACACAGCACCATTTGGCTGTC AAATAGCAACAAACCCGGTAAGAGCGATGAACTGCGCCGTAGGGAACA TGCCTATCTCCATCGACATACCGGACGCGGCCTTTACCAGGGTCGTCGA CGCGCCATCTTTAACGGACATGTCGTGTGAGGTATCAGCCTGCACCCAT TCCTCAGACTTTGGGGGCGTAGCCATCATTAAATATGCAGCCAGTAAGA AAGGCAAGTGTGCAGTGCACTCGATGACTAACGCCGTCACTATTCGGG AAGCTGAAATAGAAGTAGAAGGGAACTCTCAGTTGCAAATCTCTTTTTC GACGGCCCTAGCCAGCGCCGAATTTCGCGTACAAGTCTGTTCTACACAA GTACACTGTGCAGCCGAGTGCCATCCACCGAAAGACCATATAGTCAATT ACCCGGCGTCACACACCACCCTCGGGGTCCAAGACATTTCCGCTACGGC GATGTCATGGGTGCAGAAGATCACGGGAGGTGTGGGACTGGTTGTCGC TGTTGCAGCACTGATCCTAATCGTGGTGCTATGCGTGTCGTTTAGCAGG CACTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGG CCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCGTGGTCTT TGAATAAAGTCTGAGTGGGCGGC Chik- TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGA 378 Strain37997-E1 AATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGC (CHIKV E1 TATGGAACGAACAGCAGCCCCTGTTCTGGTTGCAGGCTCTTATCCCGCT antigen-Strain GGCCGCCTTGATCGTCCTGTGCAACTGTCTGAAACTCTTGCCATGCTGCT 37997): GTAAGACCCTGGCTTTTTTAGCCGTAATGAGCATCGGTGCCCACACTGT GAGCGCGTACGAACACGTAACAGTGATCCCGAACACGGTGGGAGTACC GTATAAGACTCTTGTCAACAGACCGGGTTACAGCCCCATGGTGTTGGAG ATGGAGCTACAATCAGTCACCTTGGAACCAACACTGTCACTTGACTACA TCACGTGCGAGTACAAAACTGTCATCCCCTCCCCGTACGTGAAGTGCTG TGGTACAGCAGAGTGCAAGGACAAGAGCCTACCAGACTACAGCTGCAA GGTCTTTACTGGAGTCTACCCATTTATGTGGGGCGGCGCCTACTGCTTTT GCGACGCCGAAAATACGCAATTGAGCGAGGCACATGTAGAGAAATCTG AATCTTGCAAAACAGAGTTTGCATCGGCCTACAGAGCCCACACCGCATC GGCGTCGGCGAAGCTCCGCGTCCTTTACCAAGGAAACAACATTACCGTA GCTGCCTACGCTAACGGTGACCATGCCGTCACAGTAAAGGACGCCAAG TTTGTCGTGGGCCCAATGTCCTCCGCCTGGACACCTTTTGACAACAAAA TCGTGGTGTACAAAGGCGACGTCTACAACATGGACTACCCACCTTTTGG CGCAGGAAGACCAGGACAATTTGGTGACATTCAAAGTCGTACACCGGA AAGTAAAGACGTTTATGCCAACACTCAGTTGGTACTACAGAGGCCAGC AGCAGGCACGGTACATGTACCATACTCTCAGGCACCATCTGGCTTCAAG TATTGGCTGAAGGAACGAGGAGCATCGCTACAGCACACGGCACCGTTC GGTTGCCAGATTGCGACAAACCCGGTAAGAGCTGTAAATTGCGCTGTG GGGAACATACCAATTTCCATCGACATACCGGATGCGGCCTTTACTAGGG TTGTCGATGCACCCTCTGTAACGGACATGTCATGCGAAGTACCAGCCTG CACTCACTCCTCCGACTTTGGGGGCGTCGCCATCATCAAATACACAGCT AGCAAGAAAGGTAAATGTGCAGTACATTCGATGACCAACGCCGTTACC ATTCGAGAAGCCGACGTAGAAGTAGAGGGGAACTCCCAGCTGCAAATA TCCTTCTCAACAGCCCTGGCAAGCGCCGAGTTTCGCGTGCAAGTGTGCT CCACACAAGTACACTGCGCAGCCGCATGCCACCCTCCAAAGGACCACA TAGTCAATTACCCAGCATCACACACCACCCTTGGGGTCCAGGATATATC CACAACGGCAATGTCTTGGGTGCAGAAGATTACGGGAGGAGTAGGATT AATTGTTGCTGTTGCTGCCTTAATTTTAATTGTGGTGCTATGCGTGTCGT TTAGCAGGCACTAATGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCT TGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACC CCCGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC chikv-Brazillian- TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGA 379 E1 (CHIKV E1 AATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGT antigen- ACGAACACGTAACAGTGATCCCGAACACGGTGGGAGTACCGTATAAGA Brazilian strain) CTCTAGTCAATAGACCGGGCTACAGTCCCATGGTATTGGAGATGGAACT ACTGTCAGTCACTTTGGAGCCAACGCTATCGCTTGATTACATCACGTGC GAGTACAAAACCGTTATCCCGTCTCCGTACGTGAAATGCTGCGGTACAG CAGAGTGCAAGGACAAAAACCTACCTGACTACAGCTGTAAGGTCTTCA CCGGCGTCTACCCATTTATGTGGGGCGGAGCCTACTGCTTCTGCGACGC TGAAAACACGCAATTGAGCGAAGCACACGTGGAGAAGTCCGAATCATG CAAAACAGAATTTGCATCAGCATACAGGGCTCATACCGCATCCGCATCA GCTAAGCTCCGCGTCCTTTACCAAGGAAATAACATCACTGTAACTGCCT ATGCTAACGGCGACCATGCCGTCACAGTTAAGGACGCCAAATTCATTGT GGGGCCAATGTCTTCAGCCTGGACACCTTTCGACAACAAAATTGTGGTG TACAAAGGTGACGTCTATAACATGGACTACCCGCCCTTTGGCGCAGGAA GACCAGGACAATTTGGCGATATCCAAAGTCGCACACCTGAGAGTAAAG ACGTCTATGCTAATACACAACTGGTACTGCAGAGACCGGCTGCGGGTAC GGTACATGTGCCATACTCTCAGGCACCATCTGGCTTTAAGTATTGGCTA AAAGAACGAGGGGCGTCGCTGCAGCACACAGCACCATTTGGCTGCCAA ATAGCAACAAACCCGGTAAGAGCGGTGAATTGCGCCGTAGGGAACATG CCCATCTCCATCGACATACCGGATGCGGCCTTCATTAGGGTCGTCGACG CGCCCTCTTTAACGGACATGTCGTGCGAGGTACCAGCCTGCACCCATTC CTCAGATTTCGGGGGCGTCGCCATTATTAAATATGCAGCCAGCAAGAAA GGCAAGTGTGCGGTGCATTCGATGACCAACGCCGTCACAATTCGGGAA GCTGAGATAGAAGTTGAAGGGAATTCTCAGCTGCAAATCTCTTTCTCGA CGGCCTTGGCCAGCGCCGAATTCCGCGTACAAGTCTGTTCTACACAAGT ACACTGTGTAGCCGAGTGCCACCCTCCGAAGGACCACATAGTCAATTAC CCGGCGTCACATACCACCCTCGGGGTCCAGGACATTTCCGCTACGGCGC TGTCATGGGTGCAGAAGATCACGGGAGGCGTGGGACTGGTTGTCGCTG TTGCAGCACTGATTCTAATCGTGGTGCTATGCGTGTCGTTCAGCAGGCA CTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCC TCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCGTGGTCTTTG AATAAAGTCTGAGTGGGCGGC ChiK.secE2 ATGGAGACCCCAGCTCAGCTTCTGTTTCTTCTCCTTCTATGGCTGCCTGA 380 HS3UPCRfree CACGACTGGACATCACCACCATCATCATAGTACAAAAGACAATTTCAAT (CHIKV secreted GTGTACAAGGCCACCCGCCCTTATTTAGCACACTGTCCAGATTGCGGTG E2 antigen): AGGGGCACTCCTGTCACTCTCCTATCGCCTTGGAGCGGATCCGGAATGA GGCGACCGATGGAACACTGAAAATCCAGGTAAGCTTGCAGATTGGCAT CAAGACTGACGATAGCCATGATTGGACCAAACTACGGTATATGGATAG CCATACACCTGCCGATGCTGAACGGGCCGGTCTGCTTGTGAGAACTAGC GCTCCATGCACCATCACGGGGACAATGGGACATTTTATCCTGGCTAGAT GCCCAAAGGGCGAAACCCTCACCGTCGGATTCACCGACTCAAGGAAAA TTTCTCACACATGTACCCATCCCTTCCACCATGAGCCACCGGTGATCGG GCGCGAACGCTTCCACAGCAGGCCTCAGCATGGAAAAGAACTGCCATG CTCGACCTATGTACAGTCCACCGCCGCTACCGCCGAAGAGATCGAAGTG CATATGCCTCCCGACACACCCGACCGAACCCTAATGACACAACAATCTG GGAATGTGAAGATTACAGTCAATGGACAGACTGTGAGGTATAAGTGTA ACTGCGGTGGCTCAAATGAGGGCCTCACCACAACGGATAAGGTGATCA ATAACTGCAAAATTGACCAGTGTCACGCGGCCGTGACCAACCATAAGA ACTGGCAGTACAACTCACCCTTAGTGCCTAGGAACGCTGAGCTGGGAG ATCGCAAGGGGAAGATACACATTCCCTTCCCGTTGGCGAATGTGACCTG CCGTGTGCCAAAAGCGAGAAATCCTACCGTAACATATGGCAAAAATCA GGTGACCATGTTGCTCTACCCGGATCACCCAACTCTGCTGAGCTATCGG AATATGGGACAAGAACCCAATTACCACGAGGAATGGGTTACGCACAAG AAAGAGGTGACCCTTACAGTCCCTACTGAAGGTCTGGAAGTGACCTGG GGCAATAACGAGCCTTATAAGTACTGGCCCCAGATGAGTACAAACGGC ACCGCCCATGGACATCCACACGAGATCATTCTGTATTACTACGAACTAT ATCCCACAATGACTGGCAAGCCTATACCAAACCCACTTCTCGGCCTTGA TAGCACATGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCT TGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCGTGG TCTTTGAATAAAGTCTGAGTGGGCGGC chikv-Brazillian- TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGA 381 E2 (CHIKV E2 AATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGA antigen- GTACCAAGGACAACTTCAATGTCTATAAAGCCACAAGACCGTACTTAGC Brazilian strain): TCACTGTCCCGACTGTGGAGAAGGGCACTCGTGCCATAGTCCCGTAGCA TTAGAACGCATCAGAAATGAAGCGACAGACGGGACGCTGAAAATCCAG GTCTCCTTGCAAATCGGAATAAAGACGGATGATAGCCACGATTGGACC AAGCTGCGTTACATGGACAACCACACGCCAGCGGACGCAGAGAGGGCG GGGCTATTTGTAAGAACATCAGCACCGTGCACGATTACTGGAACAATG GGACACTTCATCCTGACCCGATGTCCGAAAGGGGAAACTCTGACGGTG GGATTCACTGACAGTAGGAAGATCAGTCACTCATGTACGCACCCATTTC ACCACGACCCTCCTGTGATAGGCCGGGAGAAATTCCATTCCCGACCGCA GCACGGTAAAGAGCTGCCTTGCAGCACGTACGTGCAGAGCACCGCCGC AACTACCGAGGAGATAGAGGTACACATGCCCCCAGACACCCCTGATCG CACATTGATGTCACAACAGTCCGGCAACGTAAAGATCACAGTTAATGG CCAGACGGTGCGGTACAAGTGTAATTGCGGTGGCTCAAATGAAGGACT AATAACTACAGACAAAGTGATTAATAACTGCAAAGTTGATCAATGTCAT GCCGCGGTCACCAATCACAAAAAGTGGCAGTACAACTCCCCTCTGGTCC CGCGTAATGCTGAACTTGGGGACCGAAAAGGAAAAATCCACATCCCGT TTCCGCTGGCAAATGTAACATGCAGGGTGCCTAAAGCAAGGAACCCCA CCGTGACGTACGGGAAAAACCAAGTCATCATGCTACTGTATCCCGACCA CCCAACACTCCTGTCCTACCGGAACATGGGAGAAGAACCAAACTACCA AGAAGAGTGGGTGACGCATAAGAAGGAAGTCGTGCTAACCGTGCCGAC TGAAGGGCTCGAGGTCACGTGGGGTAACAACGAGCCGTATAAGTATTG GCCGCAGTTATCTACAAACGGTACAGCCCATGGCCACCCGCATGAGAT AATTCTGTATTATTATGAGCTGTACCCTACTATGACTGTAGTAGTTGTGT CAGTGGCCTCGTTCGTACTCCTGTCGATGGTGGGTGTGGCAGTGGGGAT GTGCATGTGTGCACGACGCAGATGCATCACACCGTACGAACTGACACC AGGAGCTACCGTCCCTTTCCTGCTTAGCCTAATATGCTGCATCAGAACA GCTAAAGCGTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCC CTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCGT GGTCTTTGAATAAAGTCTGAGTGGGCGGC chikv-Brazillian- TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGA 382 E2 (CHIKV E2 AATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGA antigen- GTATTAAGGACCACTTCAATGTCTATAAAGCCACAAGACCGTACCTAGC Brazilian strain): TCACTGTCCCGACTGTGGAGAAGGGCACTCGTGCCATAGTCCCGTAGCG CTAGAACGCATCAGAAACGAAGCGACAGACGGGACGTTGAAAATCCAG GTTTCCTTGCAAATCGGAATAAAGACGGATGATAGCCATGATTGGACCA AGCTGCGTTATATGGACAATCACATGCCAGCAGACGCAGAGCGGGCCG GGCTATTTGTAAGAACGTCAGCACCGTGCACGATTACTGGAACAATGG GACACTTCATTCTGGCCCGATGTCCGAAAGGAGAAACTCTGACGGTGG GGTTCACTGACGGTAGGAAGATCAGTCACTCATGTACGCACCCATTTCA CCATGACCCTCCTGTGATAGGCCGGGAAAAATTCCATTCCCGACCGCAG CACGGTAGGGAACTACCTTGCAGCACGTACGCGCAGAGCACCGCTGCA ACTGCCGAGGAGATAGAGGTACACATGCCCCCAGACACCCCAGATCGC ACATTAATGTCACAACAGTCCGGCAATGTAAAGATCACAGTCAATAGTC AGACGGTGCGGTACAAGTGCAATTGTGGTGACTCAAGTGAAGGATTAA CCACTACAGATAAAGTGATTAATAACTGCAAGGTCGATCAATGCCATGC CGCGGTCACCAATCACAAAAAATGGCAGTATAACTCCCCTCTGGTCCCG CGTAATGCTGAATTCGGGGACCGGAAAGGAAAAGTTCACATTCCATTTC CTCTGGCAAATGTGACATGCAGGGTGCCTAAAGCAAGAAACCCCACCG TGACGTACGGAAAAAACCAAGTCATCATGTTGCTGTATCCTGACCACCC AACGCTCCTGTCCTACAGGAATATGGGAGAAGAACCAAACTATCAAGA AGAGTGGGTGACGCATAAGAAGGAGATCAGGTTAACCGTGCCGACTGA GGGGCTCGAGGTCACGTGGGGTAACAATGAGCCGTACAAGTATTGGCC GCAGTTATCCACAAACGGTACAGCCCACGGCCACCCGCATGAGATAAT TCTGTATTATTATGAGCTGTACCCAACTATGACTGCGGTAGTTTTGTCAG TGGCCTCGTTCATACTCCTGTCGATGGTGGGTGTGGCAGTGGGGATGTG CATGTGTGCACGACGCAGATGCATTACACCGTACGAACTGACACCAGG AGCTACCGTCCCTTTCCTGCTTAGCCTAATATGCTGCATTAGAACAGCT AAAGCGTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTT GGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCGTGGT CTTTGAATAAAGTCTGAGTGGGCGGC Chik-Strain TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGA 383 37997-E2 AATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGC (CHIKV E2 CATATCTAGCTCATTGTCCTGACTGCGGAGAAGGGCATTCGTGCCACAG Antigen-Strain CCCTATCGCATTGGAGCGCATCAGAAATGAAGCAACGGACGGAACGCT 37997) GAAAATCCAGGTCTCTTTGCAGATCGGGATAAAGACAGATGACAGCCA CGATTGGACCAAGCTGCGCTATATGGATAGCCATACGCCAGCGGACGC GGAGCGAGCCGGATTGCTTGTAAGGACTTCAGCACCGTGCACGATCAC CGGGACCATGGGACACTTTATTCTCGCCCGATGCCCGAAAGGAGAGAC GCTGACAGTGGGATTTACGGACAGCAGAAAGATCAGCCACACATGCAC ACACCCGTTCCATCATGAACCACCTGTGATAGGTAGGGAGAGGTTCCAC TCTCGACCACAACATGGTAAAGAGTTACCTTGCAGCACGTACGTGCAGA GCACCGCTGCCACTGCTGAGGAGATAGAGGTGCATATGCCCCCAGATA CTCCTGACCGCACGCTGATGACGCAGCAGTCTGGCAACGTGAAGATCA CAGTTAATGGGCAGACGGTGCGGTACAAGTGCAACTGCGGTGGCTCAA ACGAGGGACTGACAACCACAGACAAAGTGATCAATAACTGCAAAATTG ATCAGTGCCATGCTGCAGTCACTAATCACAAGAATTGGCAATACAACTC CCCTTTAGTCCCGCGCAACGCTGAACTCGGGGACCGTAAAGGAAAGAT CCACATCCCATTCCCATTGGCAAACGTGACTTGCAGAGTGCCAAAAGCA AGAAACCCTACAGTAACTTACGGAAAAAACCAAGTCACCATGCTGCTG TATCCTGACCATCCGACACTCTTGTCTTACCGTAACATGGGACAGGAAC CAAATTACCACGAGGAGTGGGTGACACACAAGAAGGAGGTTACCTTGA CCGTGCCTACTGAGGGTCTGGAGGTCACTTGGGGCAACAACGAACCAT ACAAGTACTGGCCGCAGATGTCTACGAACGGTACTGCTCATGGTCACCC ACATGAGATAATCTTGTACTATTATGAGCTGTACCCCACTATGACTGTA GTCATTGTGTCGGTGGCCTCGTTCGTGCTTCTGTCGATGGTGGGCACAG CAGTGGGAATGTGTGTGTGCGCACGGCGCAGATGCATTACACCATATG AATTAACACCAGGAGCCACTGTTCCCTTCCTGCTCAGCCTGCTATGCTG CTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCC TCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCCCGTGGTCTTTG AATAAAGTCTGAGTGGGCGGC chikv-Brazillian- TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGA 384 E2-E1 (CHIKV AATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGA E1-E2 Antigen- GTACCAAGGACAACTTCAATGTCTATAAAGCCACAAGACCGTACTTAGC Brazilian strain): TCACTGTCCCGACTGTGGAGAAGGGCACTCGTGCCATAGTCCCGTAGCA TTAGAACGCATCAGAAATGAAGCGACAGACGGGACGCTGAAAATCCAG GTCTCCTTGCAAATCGGAATAAAGACGGATGATAGCCACGATTGGACC AAGCTGCGTTACATGGACAACCACACGCCAGCGGACGCAGAGAGGGCG GGGCTATTTGTAAGAACATCAGCACCGTGCACGATTACTGGAACAATG GGACACTTCATCCTGACCCGATGTCCGAAAGGGGAAACTCTGACGGTG GGATTCACTGACAGTAGGAAGATCAGTCACTCATGTACGCACCCATTTC ACCACGACCCTCCTGTGATAGGCCGGGAGAAATTCCATTCCCGACCGCA GCACGGTAAAGAGCTGCCTTGCAGCACGTACGTGCAGAGCACCGCCGC AACTACCGAGGAGATAGAGGTACACATGCCCCCAGACACCCCTGATCG CACATTGATGTCACAACAGTCCGGCAACGTAAAGATCACAGTTAATGG CCAGACGGTGCGGTACAAGTGTAATTGCGGTGGCTCAAATGAAGGACT AATAACTACAGACAAAGTGATTAATAACTGCAAAGTTGATCAATGTCAT GCCGCGGTCACCAATCACAAAAAGTGGCAGTACAACTCCCCTCTGGTCC CGCGTAATGCTGAACTTGGGGACCGAAAAGGAAAAATCCACATCCCGT TTCCGCTGGCAAATGTAACATGCAGGGTGCCTAAAGCAAGGAACCCCA CCGTGACGTACGGGAAAAACCAAGTCATCATGCTACTGTATCCCGACCA CCCAACACTCCTGTCCTACCGGAACATGGGAGAAGAACCAAACTACCA AGAAGAGTGGGTGACGCATAAGAAGGAAGTCGTGCTAACCGTGCCGAC TGAAGGGCTCGAGGTCACGTGGGGTAACAACGAGCCGTATAAGTATTG GCCGCAGTTATCTACAAACGGTACAGCCCATGGCCACCCGCATGAGAT AATTCTGTATTATTATGAGCTGTACCCTACTATGACTGTAGTAGTTGTGT CAGTGGCCTCGTTCGTACTCCTGTCGATGGTGGGTGTGGCAGTGGGGAT GTGCATGTGTGCACGACGCAGATGCATCACACCGTACGAACTGACACC AGGAGCTACCGTCCCTTTCCTGCTTAGCCTAATATGCTGCATCAGAACA GCTAAAGCGTACGAACACGTAACAGTGATCCCGAACACGGTGGGAGTA CCGTATAAGACTCTAGTCAATAGACCGGGCTACAGTCCCATGGTATTGG AGATGGAACTACTGTCAGTCACTTTGGAGCCAACGCTATCGCTTGATTA CATCACGTGCGAGTACAAAACCGTTATCCCGTCTCCGTACGTGAAATGC TGCGGTACAGCAGAGTGCAAGGACAAAAACCTACCTGACTACAGCTGT AAGGTCTTCACCGGCGTCTACCCATTTATGTGGGGCGGAGCCTACTGCT TCTGCGACGCTGAAAACACGCAATTGAGCGAAGCACACGTGGAGAAGT CCGAATCATGCAAAACAGAATTTGCATCAGCATACAGGGCTCATACCG CATCCGCATCAGCTAAGCTCCGCGTCCTTTACCAAGGAAATAACATCAC TGTAACTGCCTATGCTAACGGCGACCATGCCGTCACAGTTAAGGACGCC AAATTCATTGTGGGGCCAATGTCTTCAGCCTGGACACCTTTCGACAACA AAATTGTGGTGTACAAAGGTGACGTCTATAACATGGACTACCCGCCCTT TGGCGCAGGAAGACCAGGACAATTTGGCGATATCCAAAGTCGCACACC TGAGAGTAAAGACGTCTATGCTAATACACAACTGGTACTGCAGAGACC GGCTGCGGGTACGGTACATGTGCCATACTCTCAGGCACCATCTGGCTTT AAGTATTGGCTAAAAGAACGAGGGGCGTCGCTGCAGCACACAGCACCA TTTGGCTGCCAAATAGCAACAAACCCGGTAAGAGCGGTGAATTGCGCC GTAGGGAACATGCCCATCTCCATCGACATACCGGATGCGGCCTTCATTA GGGTCGTCGACGCGCCCTCTTTAACGGACATGTCGTGCGAGGTACCAGC CTGCACCCATTCCTCAGATTTCGGGGGCGTCGCCATTATTAAATATGCA GCCAGCAAGAAAGGCAAGTGTGCGGTGCATTCGATGACCAACGCCGTC ACAATTCGGGAAGCTGAGATAGAAGTTGAAGGGAATTCTCAGCTGCAA ATCTCTTTCTCGACGGCCTTGGCCAGCGCCGAATTCCGCGTACAAGTCT GTTCTACACAAGTACACTGTGTAGCCGAGTGCCACCCTCCGAAGGACCA CATAGTCAATTACCCGGCGTCACATACCACCCTCGGGGTCCAGGACATT TCCGCTACGGCGCTGTCATGGGTGCAGAAGATCACGGGAGGCGTGGGA CTGGTTGTCGCTGTTGCAGCACTGATTCTAATCGTGGTGCTATGCGTGTC GTTCAGCAGGCACTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTT GCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCC CCGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC chikv-Brazillian- TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGA 385 E2-E1 (CHIKV AATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGA E1-E2 Antigen- GTATTAAGGACCACTTCAATGTCTATAAAGCCACAAGACCGTACCTAGC Brazilian strain): TCACTGTCCCGACTGTGGAGAAGGGCACTCGTGCCATAGTCCCGTAGCG CTAGAACGCATCAGAAACGAAGCGACAGACGGGACGTTGAAAATCCAG GTTTCCTTGCAAATCGGAATAAAGACGGATGATAGCCATGATTGGACCA AGCTGCGTTATATGGACAATCACATGCCAGCAGACGCAGAGCGGGCCG GGCTATTTGTAAGAACGTCAGCACCGTGCACGATTACTGGAACAATGG GACACTTCATTCTGGCCCGATGTCCGAAAGGAGAAACTCTGACGGTGG GGTTCACTGACGGTAGGAAGATCAGTCACTCATGTACGCACCCATTTCA CCATGACCCTCCTGTGATAGGCCGGGAAAAATTCCATTCCCGACCGCAG CACGGTAGGGAACTACCTTGCAGCACGTACGCGCAGAGCACCGCTGCA ACTGCCGAGGAGATAGAGGTACACATGCCCCCAGACACCCCAGATCGC ACATTAATGTCACAACAGTCCGGCAATGTAAAGATCACAGTCAATAGTC AGACGGTGCGGTACAAGTGCAATTGTGGTGACTCAAGTGAAGGATTAA CCACTACAGATAAAGTGATTAATAACTGCAAGGTCGATCAATGCCATGC CGCGGTCACCAATCACAAAAAATGGCAGTATAACTCCCCTCTGGTCCCG CGTAATGCTGAATTCGGGGACCGGAAAGGAAAAGTTCACATTCCATTTC CTCTGGCAAATGTGACATGCAGGGTGCCTAAAGCAAGAAACCCCACCG TGACGTACGGAAAAAACCAAGTCATCATGTTGCTGTATCCTGACCACCC AACGCTCCTGTCCTACAGGAATATGGGAGAAGAACCAAACTATCAAGA AGAGTGGGTGACGCATAAGAAGGAGATCAGGTTAACCGTGCCGACTGA GGGGCTCGAGGTCACGTGGGGTAACAATGAGCCGTACAAGTATTGGCC GCAGTTATCCACAAACGGTACAGCCCACGGCCACCCGCATGAGATAAT TCTGTATTATTATGAGCTGTACCCAACTATGACTGCGGTAGTTTTGTCAG TGGCCTCGTTCATACTCCTGTCGATGGTGGGTGTGGCAGTGGGGATGTG CATGTGTGCACGACGCAGATGCATTACACCGTACGAACTGACACCAGG AGCTACCGTCCCTTTCCTGCTTAGCCTAATATGCTGCATTAGAACAGCT AAAGCGTACGAACACGTAACAGTGATCCCGAACACGGTGGGAGTACCG TATAAGACTCTAGTCAACAGACCGGGCTACAGCCCCATGGTATTGGAG ATGGAGCTTCTGTCTGTCACCTTGGAACCAACGCTATCGCTTGATTACA TCACGTGCGAGTATAAAACCGTTATCCCGTCTCCGTACGTGAAATGCTG CGGTACAGCAGAGTGTAAGGACAAGAGCCTACCTGATTACAGCTGTAA GGTCTTCACCGGCGTCTACCCATTCATGTGGGGCGGCGCCTACTGCTTC TGCGACACCGAAAATACGCAATTGAGCGAAGCACATGTGGAGAAGTCC GAATCATGCAAAACAGAATTTGCATCAGCATACAGGGCTCATACCGCA TCCGCATCAGCTAAGCTCCGCGTCCTTTACCAAGGAAATAATATCACTG TGGCTGCTTATGCAAACGGCGACCATGCCGTCACAGTTAAGGACGCTAA ATTCATAGTGGGGCCAATGTCTTCAGCCTGGACACCTTTCGACAATAAA ATCGTGGTGTACAAAGGCGACGTCTACAACATGGACTACCCGCCCTTCG GCGCAGGAAGACCAGGACAATTTGGCGACATCCAAAGTCGCACGCCTG AGAGCGAAGACGTCTATGCTAATACACAACTGGTACTGCAGAGACCGT CCGCGGGTACGGTGCACGTGCCGTACTCTCAGGCACCATCTGGCTTCAA GTATTGGCTAAAAGAACGAGGGGCGTCGCTGCAGCACACAGCACCATT TGGCTGTCAAATAGCAACAAACCCGGTAAGAGCGATGAACTGCGCCGT AGGGAACATGCCTATCTCCATCGACATACCGGACGCGGCCTTTACCAGG GTCGTCGACGCGCCATCTTTAACGGACATGTCGTGTGAGGTATCAGCCT GCACCCATTCCTCAGACTTTGGGGGCGTAGCCATCATTAAATATGCAGC CAGTAAGAAAGGCAAGTGTGCAGTGCACTCGATGACTAACGCCGTCAC TATTCGGGAAGCTGAAATAGAAGTAGAAGGGAACTCTCAGTTGCAAAT CTCTTTTTCGACGGCCCTAGCCAGCGCCGAATTTCGCGTACAAGTCTGTT CTACACAAGTACACTGTGCAGCCGAGTGCCATCCACCGAAAGACCATA TAGTCAATTACCCGGCGTCACACACCACCCTCGGGGTCCAAGACATTTC CGCTACGGCGATGTCATGGGTGCAGAAGATCACGGGAGGTGTGGGACT GGTTGTCGCTGTTGCAGCACTGATCCTAATCGTGGTGCTATGCGTGTCG TTTAGCAGGCACATGAGTATTAAGGACCACTTCAATGTCTATAAAGCCA CAAGACCGTACCTAGCTCACTGTCCCGACTGTGGAGAAGGGCACTCGTG CCATAGTCCCGTAGCGCTAGAACGCATCAGAAACGAAGCGACAGACGG GACGTTGAAAATCCAGGTTTCCTTGCAAATCGGAATAAAGACGGATGA TAGCCATGATTGGACCAAGCTGCGTTATATGGACAATCACATGCCAGCA GACGCAGAGCGGGCCGGGCTATTTGTAAGAACGTCAGCACCGTGCACG ATTACTGGAACAATGGGACACTTCATTCTGGCCCGATGTCCGAAAGGAG AAACTCTGACGGTGGGGTTCACTGACGGTAGGAAGATCAGTCACTCAT GTACGCACCCATTTCACCATGACCCTCCTGTGATAGGCCGGGAAAAATT CCATTCCCGACCGCAGCACGGTAGGGAACTACCTTGCAGCACGTACGC GCAGAGCACCGCTGCAACTGCCGAGGAGATAGAGGTACACATGCCCCC AGACACCCCAGATCGCACATTAATGTCACAACAGTCCGGCAATGTAAA GATCACAGTCAATAGTCAGACGGTGCGGTACAAGTGCAATTGTGGTGA CTCAAGTGAAGGATTAACCACTACAGATAAAGTGATTAATAACTGCAA GGTCGATCAATGCCATGCCGCGGTCACCAATCACAAAAAATGGCAGTA TAACTCCCCTCTGGTCCCGCGTAATGCTGAATTCGGGGACCGGAAAGGA AAAGTTCACATTCCATTTCCTCTGGCAAATGTGACATGCAGGGTGCCTA AAGCAAGAAACCCCACCGTGACGTACGGAAAAAACCAAGTCATCATGT TGCTGTATCCTGACCACCCAACGCTCCTGTCCTACAGGAATATGGGAGA AGAACCAAACTATCAAGAAGAGTGGGTGACGCATAAGAAGGAGATCA GGTTAACCGTGCCGACTGAGGGGCTCGAGGTCACGTGGGGTAACAATG AGCCGTACAAGTATTGGCCGCAGTTATCCACAAACGGTACAGCCCACG GCCACCCGCATGAGATAATTCTGTATTATTATGAGCTGTACCCAACTAT GACTGCGGTAGTTTTGTCAGTGGCCTCGTTCATACTCCTGTCGATGGTG GGTGTGGCAGTGGGGATGTGCATGTGTGCACGACGCAGATGCATTACA CCGTACGAACTGACACCAGGAGCTACCGTCCCTTTCCTGCTTAGCCTAA TATGCTGCATTAGAACAGCTAAAGCGTACGAACACGTAACAGTGATCC CGAACACGGTGGGAGTACCGTATAAGACTCTAGTCAACAGACCGGGCT ACAGCCCCATGGTATTGGAGATGGAGCTTCTGTCTGTCACCTTGGAACC AACGCTATCGCTTGATTACATCACGTGCGAGTATAAAACCGTTATCCCG TCTCCGTACGTGAAATGCTGCGGTACAGCAGAGTGTAAGGACAAGAGC CTACCTGATTACAGCTGTAAGGTCTTCACCGGCGTCTACCCATTCATGT GGGGCGGCGCCTACTGCTTCTGCGACACCGAAAATACGCAATTGAGCG AAGCACATGTGGAGAAGTCCGAATCATGCAAAACAGAATTTGCATCAG CATACAGGGCTCATACCGCATCCGCATCAGCTAAGCTCCGCGTCCTTTA CCAAGGAAATAATATCACTGTGGCTGCTTATGCAAACGGCGACCATGCC GTCACAGTTAAGGACGCTAAATTCATAGTGGGGCCAATGTCTTCAGCCT GGACACCTTTCGACAATAAAATCGTGGTGTACAAAGGCGACGTCTACA ACATGGACTACCCGCCCTTCGGCGCAGGAAGACCAGGACAATTTGGCG ACATCCAAAGTCGCACGCCTGAGAGCGAAGACGTCTATGCTAATACAC AACTGGTACTGCAGAGACCGTCCGCGGGTACGGTGCACGTGCCGTACTC TCAGGCACCATCTGGCTTCAAGTATTGGCTAAAAGAACGAGGGGCGTC GCTGCAGCACACAGCACCATTTGGCTGTCAAATAGCAACAAACCCGGT AAGAGCGATGAACTGCGCCGTAGGGAACATGCCTATCTCCATCGACAT ACCGGACGCGGCCTTTACCAGGGTCGTCGACGCGCCATCTTTAACGGAC ATGTCGTGTGAGGTATCAGCCTGCACCCATTCCTCAGACTTTGGGGGCG TAGCCATCATTAAATATGCAGCCAGTAAGAAAGGCAAGTGTGCAGTGC ACTCGATGACTAACGCCGTCACTATTCGGGAAGCTGAAATAGAAGTAG AAGGGAACTCTCAGTTGCAAATCTCTTTTTCGACGGCCCTAGCCAGCGC CGAATTTCGCGTACAAGTCTGTTCTACACAAGTACACTGTGCAGCCGAG TGCCATCCACCGAAAGACCATATAGTCAATTACCCGGCGTCACACACCA CCCTCGGGGTCCAAGACATTTCCGCTACGGCGATGTCATGGGTGCAGAA GATCACGGGAGGTGTGGGACTGGTTGTCGCTGTTGCAGCACTGATCCTA ATCGTGGTGCTATGCGTGTCGTTTAGCAGGCACTGATAATAGGCTGGAG CCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTC CCCTTCCTGCACCCGTACCCCCGTGGTCTTTGAATAAAGTCTGAGTGGG CGGC Chik-Strain TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGA 386 37997-E2-E1 AATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGC (CHIKV E1-E2 CATATCTAGCTCATTGTCCTGACTGCGGAGAAGGGCATTCGTGCCACAG Antigen-Strain CCCTATCGCATTGGAGCGCATCAGAAATGAAGCAACGGACGGAACGCT 37997): GAAAATCCAGGTCTCTTTGCAGATCGGGATAAAGACAGATGACAGCCA CGATTGGACCAAGCTGCGCTATATGGATAGCCATACGCCAGCGGACGC GGAGCGAGCCGGATTGCTTGTAAGGACTTCAGCACCGTGCACGATCAC CGGGACCATGGGACACTTTATTCTCGCCCGATGCCCGAAAGGAGAGAC GCTGACAGTGGGATTTACGGACAGCAGAAAGATCAGCCACACATGCAC ACACCCGTTCCATCATGAACCACCTGTGATAGGTAGGGAGAGGTTCCAC TCTCGACCACAACATGGTAAAGAGTTACCTTGCAGCACGTACGTGCAGA GCACCGCTGCCACTGCTGAGGAGATAGAGGTGCATATGCCCCCAGATA CTCCTGACCGCACGCTGATGACGCAGCAGTCTGGCAACGTGAAGATCA CAGTTAATGGGCAGACGGTGCGGTACAAGTGCAACTGCGGTGGCTCAA ACGAGGGACTGACAACCACAGACAAAGTGATCAATAACTGCAAAATTG ATCAGTGCCATGCTGCAGTCACTAATCACAAGAATTGGCAATACAACTC CCCTTTAGTCCCGCGCAACGCTGAACTCGGGGACCGTAAAGGAAAGAT CCACATCCCATTCCCATTGGCAAACGTGACTTGCAGAGTGCCAAAAGCA AGAAACCCTACAGTAACTTACGGAAAAAACCAAGTCACCATGCTGCTG TATCCTGACCATCCGACACTCTTGTCTTACCGTAACATGGGACAGGAAC CAAATTACCACGAGGAGTGGGTGACACACAAGAAGGAGGTTACCTTGA CCGTGCCTACTGAGGGTCTGGAGGTCACTTGGGGCAACAACGAACCAT ACAAGTACTGGCCGCAGATGTCTACGAACGGTACTGCTCATGGTCACCC ACATGAGATAATCTTGTACTATTATGAGCTGTACCCCACTATGACTGTA GTCATTGTGTCGGTGGCCTCGTTCGTGCTTCTGTCGATGGTGGGCACAG CAGTGGGAATGTGTGTGTGCGCACGGCGCAGATGCATTACACCATATG AATTAACACCAGGAGCCACTGTTCCCTTCCTGCTCAGCCTGCTATGCTG CCTATGGAACGAACAGCAGCCCCTGTTCTGGTTGCAGGCTCTTATCCCG CTGGCCGCCTTGATCGTCCTGTGCAACTGTCTGAAACTCTTGCCATGCTG CTGTAAGACCCTGGCTTTTTTAGCCGTAATGAGCATCGGTGCCCACACT GTGAGCGCGTACGAACACGTAACAGTGATCCCGAACACGGTGGGAGTA CCGTATAAGACTCTTGTCAACAGACCGGGTTACAGCCCCATGGTGTTGG AGATGGAGCTACAATCAGTCACCTTGGAACCAACACTGTCACTTGACTA CATCACGTGCGAGTACAAAACTGTCATCCCCTCCCCGTACGTGAAGTGC TGTGGTACAGCAGAGTGCAAGGACAAGAGCCTACCAGACTACAGCTGC AAGGTCTTTACTGGAGTCTACCCATTTATGTGGGGCGGCGCCTACTGCT TTTGCGACGCCGAAAATACGCAATTGAGCGAGGCACATGTAGAGAAAT CTGAATCTTGCAAAACAGAGTTTGCATCGGCCTACAGAGCCCACACCGC ATCGGCGTCGGCGAAGCTCCGCGTCCTTTACCAAGGAAACAACATTACC GTAGCTGCCTACGCTAACGGTGACCATGCCGTCACAGTAAAGGACGCC AAGTTTGTCGTGGGCCCAATGTCCTCCGCCTGGACACCTTTTGACAACA AAATCGTGGTGTACAAAGGCGACGTCTACAACATGGACTACCCACCTTT TGGCGCAGGAAGACCAGGACAATTTGGTGACATTCAAAGTCGTACACC GGAAAGTAAAGACGTTTATGCCAACACTCAGTTGGTACTACAGAGGCC AGCAGCAGGCACGGTACATGTACCATACTCTCAGGCACCATCTGGCTTC AAGTATTGGCTGAAGGAACGAGGAGCATCGCTACAGCACACGGCACCG TTCGGTTGCCAGATTGCGACAAACCCGGTAAGAGCTGTAAATTGCGCTG TGGGGAACATACCAATTTCCATCGACATACCGGATGCGGCCTTTACTAG GGTTGTCGATGCACCCTCTGTAACGGACATGTCATGCGAAGTACCAGCC TGCACTCACTCCTCCGACTTTGGGGGCGTCGCCATCATCAAATACACAG CTAGCAAGAAAGGTAAATGTGCAGTACATTCGATGACCAACGCCGTTA CCATTCGAGAAGCCGACGTAGAAGTAGAGGGGAACTCCCAGCTGCAAA TATCCTTCTCAACAGCCCTGGCAAGCGCCGAGTTTCGCGTGCAAGTGTG CTCCACACAAGTACACTGCGCAGCCGCATGCCACCCTCCAAAGGACCA CATAGTCAATTACCCAGCATCACACACCACCCTTGGGGTCCAGGATATA TCCACAACGGCAATGTCTTGGGTGCAGAAGATTACGGGAGGAGTAGGA TTAATTGTTGCTGTTGCTGCCTTAATTTTAATTGTGGTGCTATGCGTGTC GTTTAGCAGGCACTAATGATAATAGGCTGGAGCCTCGGTGGCCATGCTT CTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTA CCCCCGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC Chik.C-E3-E2- TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGA 387 6K- AATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGG E1_HS3UPCRfree AGTTTATCCCTACGCAGACGTTCTATAATCGGAGGTACCAGCCCAGGCC (C-E3-E2-6K- TTGGGCCCCCCGCCCTACAATCCAAGTGATAAGACCACGTCCCAGGCCG E1 Antigen) CAGAGACAAGCCGGCCAATTGGCGCAACTCATCAGCGCAGTTAACAAG TTGACCATGCGAGCGGTTCCTCAGCAGAAGCCGAGGCGGAACCGGAAG AATAAGAAACAACGCCAAAAGAAACAGGCGCCGCAGAACGACCCTAA ACAGAAGAAACAACCTCCCCAGAAAAAGCCAGCTCAGAAGAAGAAGA AGCCTGGACGCCGTGAAAGAATGTGCATGAAAATCGAAAATGATTGCA TCTTTGAGGTGAAGCACGAGGGCAAAGTGATGGGGTACGCATGCCTGG TGGGCGATAAGGTCATGAAGCCAGCACATGTGAAGGGGACAATCGATA ATGCTGATCTGGCCAAGCTAGCTTTTAAACGTAGCTCCAAATACGATCT TGAGTGTGCCCAGATACCTGTGCACATGAAATCTGATGCAAGCAAGTTC ACACACGAGAAGCCTGAGGGCTATTATAACTGGCATCATGGTGCGGTTC AGTACTCCGGCGGCCGATTTACCATTCCTACAGGGGCAGGAAAGCCGG GCGATTCGGGGAGACCCATTTTCGACAACAAAGGCCGCGTGGTAGCTA TCGTGCTCGGTGGGGCTAATGAGGGTGCACGTACTGCACTTAGCGTGGT TACCTGGAATAAGGACATTGTCACAAAGATTACACCGGAGGGAGCAGA GGAATGGAGCCTGGCACTGCCCGTTCTGTGCCTGCTGGCCAACACCACT TTCCCATGTAGTCAACCCCCTTGCACTCCCTGCTGCTATGAGAAAGAGC CTGAGAGCACGTTACGTATGCTGGAAGATAATGTCATGAGGCCCGGGT ACTATCAACTGCTCAAGGCTAGTCTGACATGCTCGCCCCACAGGCAGCG CAGGTCCACGAAAGATAACTTCAACGTTTACAAGGCTACTAGGCCTTAT TTGGCCCACTGTCCCGATTGCGGAGAGGGACATTCTTGTCATAGTCCTA TTGCCTTGGAGCGAATCCGCAACGAGGCCACTGATGGAACCCTTAAGAT TCAAGTATCTTTGCAGATTGGCATTAAGACAGATGATTCCCATGACTGG ACAAAGCTTCGGTACATGGACTCACACACGCCTGCAGATGCTGAAAGG GCAGGGCTCTTGGTCAGGACCTCGGCCCCTTGTACAATTACCGGGACCA TGGGCCACTTCATCCTTGCACGCTGCCCTAAGGGGGAGACGCTGACGGT GGGCTTTACTGACTCGCGTAAGATCTCACACACATGTACACACCCTTTC CACCACGAACCTCCAGTCATAGGGAGAGAGAGATTTCACTCTCGCCCAC AGCATGGCAAAGAGCTGCCATGCTCCACATATGTCCAGAGCACTGCTGC TACCGCTGAAGAAATTGAGGTTCACATGCCACCCGATACACCAGACCGT ACTCTGATGACCCAACAGAGCGGCAACGTGAAGATTACCGTAAATGGA CAGACCGTGAGATATAAGTGCAACTGTGGTGGCTCCAATGAGGGCTTA ACAACAACGGATAAGGTGATTAACAATTGCAAAATAGATCAGTGCCAT GCCGCAGTGACCAATCACAAGAATTGGCAATACAACTCACCCCTAGTG CCGAGGAACGCAGAACTAGGCGACAGGAAAGGGAAAATCCATATACCC TTCCCCCTAGCAAATGTGACCTGCCGAGTGCCCAAGGCCAGAAACCCCA CGGTTACTTACGGCAAGAACCAGGTGACGATGCTTTTGTACCCAGACCA TCCCACCTTGCTCTCTTATAGAAACATGGGACAGGAGCCTAACTATCAT GAGGAGTGGGTGACACACAAGAAAGAAGTCACCCTTACCGTGCCTACC GAAGGGCTTGAAGTCACCTGGGGCAACAACGAGCCTTACAAGTATTGG CCACAGATGTCCACAAACGGAACAGCCCACGGCCACCCGCACGAGATC ATACTGTATTACTATGAGCTTTATCCCACAATGACTGTCGTAATTGTGA GCGTTGCCAGCTTCGTGTTGCTTTCAATGGTTGGCACTGCCGTGGGGAT GTGCGTGTGTGCTAGGCGCCGCTGTATAACTCCTTATGAACTAACTCCA GGCGCCACCGTTCCTTTCCTGCTCTCACTACTGTGTTGTGTGCGCACAAC AAAGGCTGCCACCTACTACGAAGCCGCCGCCTACTTATGGAATGAACA GCAGCCTCTCTTTTGGTTACAGGCGCTGATTCCTCTTGCTGCCCTGATCG TGCTATGCAACTGCCTCAAGCTGCTGCCCTGTTGTTGCAAGACCCTAGC TTTTCTCGCCGTGATGAGCATCGGGGCACATACAGTGTCCGCCTATGAG CACGTCACCGTTATCCCGAACACCGTCGGTGTGCCATATAAGACGTTAG TCAATCGACCCGGCTACTCTCCAATGGTGCTGGAAATGGAACTCCAGAG TGTGACACTGGAGCCAACCTTATCCCTCGATTATATTACCTGCGAATAC AAGACCGTCATCCCTTCACCCTATGTCAAGTGCTGTGGGACCGCTGAAT GCAAAGACAAGAGCTTGCCTGATTACAGTTGCAAGGTCTTCACAGGTGT GTACCCCTTCATGTGGGGGGGAGCTTATTGCTTTTGTGATGCTGAGAAC ACCCAACTGAGCGAGGCTCACGTCGAGAAATCTGAGTCTTGCAAGACC GAGTTTGCCTCAGCTTACAGGGCCCACACGGCCAGCGCATCCGCCAAAT TGAGGGTACTCTACCAGGGTAATAATATCACCGTTGCCGCATATGCAAA CGGCGATCACGCCGTGACTGTCAAGGATGCCAAGTTCGTTGTGGGCCCC ATGTCTAGCGCTTGGACACCGTTCGATAATAAGATCGTCGTGTACAAAG GGGACGTGTATAATATGGACTACCCACCTTTCGGGGCCGGCCGACCGG GACAGTTCGGGGATATTCAGAGCCGCACACCCGAATCTAAAGATGTTTA CGCCAATACTCAGCTCGTCCTGCAGAGGCCCGCCGCTGGTACAGTTCAC GTTCCTTACTCACAGGCACCCTCTGGGTTTAAGTATTGGCTGAAAGAAC GAGGTGCCAGCTTGCAGCATACAGCGCCTTTCGGATGCCAGATTGCCAC TAACCCCGTACGGGCTGTCAACTGCGCGGTCGGCAATATTCCCATTAGC ATTGATATCCCGGACGCAGCTTTCACCAGGGTTGTGGACGCCCCGAGCG TCACCGACATGAGTTGTGAGGTGCCAGCCTGCACGCATAGCAGTGATTT CGGCGGCGTCGCCATCATTAAATATACCGCAAGCAAGAAAGGCAAGTG CGCCGTCCACTCGATGACTAACGCCGTCACAATTCGGGAAGCCGATGTT GAGGTCGAAGGCAACTCCCAGCTGCAGATCAGCTTCTCTACTGCTCTTG CAAGCGCCGAGTTTCGAGTCCAGGTCTGCAGTACGCAGGTGCATTGTGC AGCTGCCTGCCATCCACCCAAAGATCATATTGTGAATTATCCGGCGTCA CATACCACACTGGGGGTCCAGGATATTAGTACAACGGCGATGTCCTGG GTGCAGAAAATTACGGGAGGAGTGGGCTTAATTGTTGCCGTGGCGGCC CTGATCCTGATCGTTGTGCTGTGTGTTAGCTTCTCTAGGCATGACTATAA AGATGACGATGACAAATGATAATAGGCTGGAGCCTCGGTGGCCATGCT TCTTGCCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGT ACCCCCGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC CHIKV C-E3- TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGA 388 E2-6K-E1 AATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGG AGTTTATCCCTACGCAGACGTTCTATAATCGGAGGTACCAGCCCAGGCC TTGGGCCCCCCGCCCTACAATCCAAGTGATAAGACCACGTCCCAGGCCG CAGAGACAAGCCGGCCAATTGGCGCAACTCATCAGCGCAGTTAACAAG TTGACCATGCGAGCGGTTCCTCAGCAGAAGCCGAGGCGGAACCGGAAG AATAAGAAACAACGCCAAAAGAAACAGGCGCCGCAGAACGACCCTAA ACAGAAGAAACAACCTCCCCAGAAAAAGCCAGCTCAGAAGAAGAAGA AGCCTGGACGCCGTGAAAGAATGTGCATGAAAATCGAAAATGATTGCA TCTTTGAGGTGAAGCACGAGGGCAAAGTGATGGGGTACGCATGCCTGG TGGGCGATAAGGTCATGAAGCCAGCACATGTGAAGGGGACAATCGATA ATGCTGATCTGGCCAAGCTAGCTTTTAAACGTAGCTCCAAATACGATCT TGAGTGTGCCCAGATACCTGTGCACATGAAATCTGATGCAAGCAAGTTC ACACACGAGAAGCCTGAGGGCTATTATAACTGGCATCATGGTGCGGTTC AGTACTCCGGCGGCCGATTTACCATTCCTACAGGGGCAGGAAAGCCGG GCGATTCGGGGAGACCCATTTTCGACAACAAAGGCCGCGTGGTAGCTA TCGTGCTCGGTGGGGCTAATGAGGGTGCACGTACTGCACTTAGCGTGGT TACCTGGAATAAGGACATTGTCACAAAGATTACACCGGAGGGAGCAGA GGAATGGAGCCTGGCACTGCCCGTTCTGTGCCTGCTGGCCAACACCACT TTCCCATGTAGTCAACCCCCTTGCACTCCCTGCTGCTATGAGAAAGAGC CTGAGAGCACGTTACGTATGCTGGAAGATAATGTCATGAGGCCCGGGT ACTATCAACTGCTCAAGGCTAGTCTGACATGCTCGCCCCACAGGCAGCG CAGGTCCACGAAAGATAACTTCAACGTTTACAAGGCTACTAGGCCTTAT TTGGCCCACTGTCCCGATTGCGGAGAGGGACATTCTTGTCATAGTCCTA TTGCCTTGGAGCGAATCCGCAACGAGGCCACTGATGGAACCCTTAAGAT TCAAGTATCTTTGCAGATTGGCATTAAGACAGATGATTCCCATGACTGG ACAAAGCTTCGGTACATGGACTCACACACGCCTGCAGATGCTGAAAGG GCAGGGCTCTTGGTCAGGACCTCGGCCCCTTGTACAATTACCGGGACCA TGGGCCACTTCATCCTTGCACGCTGCCCTAAGGGGGAGACGCTGACGGT GGGCTTTACTGACTCGCGTAAGATCTCACACACATGTACACACCCTTTC CACCACGAACCTCCAGTCATAGGGAGAGAGAGATTTCACTCTCGCCCAC AGCATGGCAAAGAGCTGCCATGCTCCACATATGTCCAGAGCACTGCTGC TACCGCTGAAGAAATTGAGGTTCACATGCCACCCGATACACCAGACCGT ACTCTGATGACCCAACAGAGCGGCAACGTGAAGATTACCGTAAATGGA CAGACCGTGAGATATAAGTGCAACTGTGGTGGCTCCAATGAGGGCTTA ACAACAACGGATAAGGTGATTAACAATTGCAAAATAGATCAGTGCCAT GCCGCAGTGACCAATCACAAGAATTGGCAATACAACTCACCCCTAGTG CCGAGGAACGCAGAACTAGGCGACAGGAAAGGGAAAATCCATATACCC TTCCCCCTAGCAAATGTGACCTGCCGAGTGCCCAAGGCCAGAAACCCCA CGGTTACTTACGGCAAGAACCAGGTGACGATGCTTTTGTACCCAGACCA TCCCACCTTGCTCTCTTATAGAAACATGGGACAGGAGCCTAACTATCAT GAGGAGTGGGTGACACACAAGAAAGAAGTCACCCTTACCGTGCCTACC GAAGGGCTTGAAGTCACCTGGGGCAACAACGAGCCTTACAAGTATTGG CCACAGATGTCCACAAACGGAACAGCCCACGGCCACCCGCACGAGATC ATACTGTATTACTATGAGCTTTATCCCACAATGACTGTCGTAATTGTGA GCGTTGCCAGCTTCGTGTTGCTTTCAATGGTTGGCACTGCCGTGGGGAT GTGCGTGTGTGCTAGGCGCCGCTGTATAACTCCTTATGAACTAACTCCA GGCGCCACCGTTCCTTTCCTGCTCTCACTACTGTGTTGTGTGCGCACAAC AAAGGCTGCCACCTACTACGAAGCCGCCGCCTACTTATGGAATGAACA GCAGCCTCTCTTTTGGTTACAGGCGCTGATTCCTCTTGCTGCCCTGATCG TGCTATGCAACTGCCTCAAGCTGCTGCCCTGTTGTTGCAAGACCCTAGC TTTTCTCGCCGTGATGAGCATCGGGGCACATACAGTGTCCGCCTATGAG CACGTCACCGTTATCCCGAACACCGTCGGTGTGCCATATAAGACGTTAG TCAATCGACCCGGCTACTCTCCAATGGTGCTGGAAATGGAACTCCAGAG TGTGACACTGGAGCCAACCTTATCCCTCGATTATATTACCTGCGAATAC AAGACCGTCATCCCTTCACCCTATGTCAAGTGCTGTGGGACCGCTGAAT GCAAAGACAAGAGCTTGCCTGATTACAGTTGCAAGGTCTTCACAGGTGT GTACCCCTTCATGTGGGGGGGAGCTTATTGCTTTTGTGATGCTGAGAAC ACCCAACTGAGCGAGGCTCACGTCGAGAAATCTGAGTCTTGCAAGACC GAGTTTGCCTCAGCTTACAGGGCCCACACGGCCAGCGCATCCGCCAAAT TGAGGGTACTCTACCAGGGTAATAATATCACCGTTGCCGCATATGCAAA CGGCGATCACGCCGTGACTGTCAAGGATGCCAAGTTCGTTGTGGGCCCC ATGTCTAGCGCTTGGACACCGTTCGATAATAAGATCGTCGTGTACAAAG GGGACGTGTATAATATGGACTACCCACCTTTCGGGGCCGGCCGACCGG GACAGTTCGGGGATATTCAGAGCCGCACACCCGAATCTAAAGATGTTTA CGCCAATACTCAGCTCGTCCTGCAGAGGCCCGCCGCTGGTACAGTTCAC GTTCCTTACTCACAGGCACCCTCTGGGTTTAAGTATTGGCTGAAAGAAC GAGGTGCCAGCTTGCAGCATACAGCGCCTTTCGGATGCCAGATTGCCAC TAACCCCGTACGGGCTGTCAACTGCGCGGTCGGCAATATTCCCATTAGC ATTGATATCCCGGACGCAGCTTTCACCAGGGTTGTGGACGCCCCGAGCG TCACCGACATGAGTTGTGAGGTGCCAGCCTGCACGCATAGCAGTGATTT CGGCGGCGTCGCCATCATTAAATATACCGCAAGCAAGAAAGGCAAGTG CGCCGTCCACTCGATGACTAACGCCGTCACAATTCGGGAAGCCGATGTT GAGGTCGAAGGCAACTCCCAGCTGCAGATCAGCTTCTCTACTGCTCTTG CAAGCGCCGAGTTTCGAGTCCAGGTCTGCAGTACGCAGGTGCATTGTGC AGCTGCCTGCCATCCACCCAAAGATCATATTGTGAATTATCCGGCGTCA CATACCACACTGGGGGTCCAGGATATTAGTACAACGGCGATGTCCTGG GTGCAGAAAATTACGGGAGGAGTGGGCTTAATTGTTGCCGTGGCGGCC CTGATCCTGATCGTTGTGCTGTGTGTTAGCTTCTCTAGGCATTGATAATA GGCTGGAGCCTCGGTGGCCATGCTTCTTGCCCCTTGGGCCTCCCCCCAG CCCCTCCTCCCCTTCCTGCACCCGTACCCCCGTGGTCTTTGAATAAAGTC TGAGTGGGCGGC CHIKV mRNA Sequences ChiK.secE1 UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGG 389 HS3UPCRfree GAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCA (CHIKV secreted UGGAGACACCUGCACAGCUGUUGUUUCUGCUGCUGCUUUGGUUGCCC E1 antigen) GAUACCACCGGUGACUACAAAGACGACGACGAUAAAUACGAGCACGU GACGGUAAUACCAAACACUGUGGGGGUGCCAUACAAGACCCUGGUAA AUCGCCCAGGCUACUCUCCCAUGGUGCUGGAGAUGGAGCUCCAGUCU GUGACCUUAGAGCCAACCCUCUCACUCGACUAUAUCACCUGUGAAUA CAAAACAGUGAUCCCAUCCCCCUACGUGAAAUGUUGCGGAACUGCAG AGUGUAAGGAUAAGAGUCUGCCCGAUUACAGCUGCAAGGUGUUUACA GGCGUGUAUCCAUUUAUGUGGGGAGGAGCCUACUGUUUUUGCGAUGC CGAAAAUACUCAGCUGUCUGAAGCCCAUGUGGAGAAGAGUGAAAGUU GCAAGACCGAAUUUGCUAGUGCCUACAGGGCACACACCGCUUCUGCC UCCGCUAAACUCCGAGUCCUUUACCAGGGCAAUAAUAUUACGGUCGC UGCCUACGCUAACGGGGACCACGCUGUGACAGUCAAGGACGCCAAAU UCGUAGUGGGCCCAAUGAGCUCCGCCUGGACUCCCUUCGACAACAAA AUCGUCGUGUAUAAAGGCGACGUGUACAAUAUGGACUACCCACCCUU CGGGGCUGGAAGACCGGGCCAGUUUGGAGAUAUCCAAUCAAGGACAC CCGAGUCAAAGGACGUGUACGCCAAUACGCAGCUGGUGCUGCAGAGA CCCGCCGCUGGUACCGUGCAUGUGCCUUAUUCCCAAGCUCCAUCUGG CUUCAAGUACUGGUUGAAAGAGCGCGGUGCUUCGCUGCAGCAUACAG CACCGUUCGGAUGUCAGAUAGCAACCAACCCUGUACGGGCUGUCAAC UGUGCCGUGGGAAAUAUACCUAUUUCCAUCGACAUUCCGGACGCAGC UUUCACACGUGUCGUUGAUGCCCCCUCAGUGACUGACAUGUCAUGUG AGGUGCCUGCUUGCACCCACAGCAGCGAUUUUGGCGGAGUGGCCAUA AUCAAGUACACCGCCUCCAAAAAAGGAAAGUGUGCCGUACACUCUAU GACCAACGCCGUCACAAUCAGAGAAGCCGACGUUGAAGUAGAGGGAA AUUCACAGCUGCAAAUCAGCUUCAGCACCGCUCUUGCCUCUGCUGAG UUUAGGGUUCAGGUUUGCAGUACUCAGGUGCACUGUGCAGCCGCUUG CCAUCCCCCCAAGGAUCAUAUCGUGAAUUAUCCUGCAUCCCACACCAC ACUGGGAGUCCAGGAUAUCUCAACAACUGCAAUGUCUUGGGUGCAGA AGAUCACCUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCC CCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCC CGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC Chik- UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGG 390 Strain37997-E1 GAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCA (CHIKV E1 UGUACGAACACGUAACAGUGAUCCCGAACACGGUGGGAGUACCGUAU antigen-Strain AAGACUCUAGUCAACAGACCGGGCUACAGCCCCAUGGUAUUGGAGAU 37997): GGAGCUUCUGUCUGUCACCUUGGAACCAACGCUAUCGCUUGAUUACA UCACGUGCGAGUAUAAAACCGUUAUCCCGUCUCCGUACGUGAAAUGC UGCGGUACAGCAGAGUGUAAGGACAAGAGCCUACCUGAUUACAGCUG UAAGGUCUUCACCGGCGUCUACCCAUUCAUGUGGGGCGGCGCCUACU GCUUCUGCGACACCGAAAAUACGCAAUUGAGCGAAGCACAUGUGGAG AAGUCCGAAUCAUGCAAAACAGAAUUUGCAUCAGCAUACAGGGCUCA UACCGCAUCCGCAUCAGCUAAGCUCCGCGUCCUUUACCAAGGAAAUA AUAUCACUGUGGCUGCUUAUGCAAACGGCGACCAUGCCGUCACAGUU AAGGACGCUAAAUUCAUAGUGGGGCCAAUGUCUUCAGCCUGGACACC UUUCGACAAUAAAAUCGUGGUGUACAAAGGCGACGUCUACAACAUGG ACUACCCGCCCUUCGGCGCAGGAAGACCAGGACAAUUUGGCGACAUC CAAAGUCGCACGCCUGAGAGCGAAGACGUCUAUGCUAAUACACAACU GGUACUGCAGAGACCGUCCGCGGGUACGGUGCACGUGCCGUACUCUC AGGCACCAUCUGGCUUCAAGUAUUGGCUAAAAGAACGAGGGGCGUCG CUGCAGCACACAGCACCAUUUGGCUGUCAAAUAGCAACAAACCCGGU AAGAGCGAUGAACUGCGCCGUAGGGAACAUGCCUAUCUCCAUCGACA UACCGGACGCGGCCUUUACCAGGGUCGUCGACGCGCCAUCUUUAACG GACAUGUCGUGUGAGGUAUCAGCCUGCACCCAUUCCUCAGACUUUGG GGGCGUAGCCAUCAUUAAAUAUGCAGCCAGUAAGAAAGGCAAGUGUG CAGUGCACUCGAUGACUAACGCCGUCACUAUUCGGGAAGCUGAAAUA GAAGUAGAAGGGAACUCUCAGUUGCAAAUCUCUUUUUCGACGGCCCU AGCCAGCGCCGAAUUUCGCGUACAAGUCUGUUCUACACAAGUACACU GUGCAGCCGAGUGCCAUCCACCGAAAGACCAUAUAGUCAAUUACCCG GCGUCACACACCACCCUCGGGGUCCAAGACAUUUCCGCUACGGCGAU GUCAUGGGUGCAGAAGAUCACGGGAGGUGUGGGACUGGUUGUCGCUG UUGCAGCACUGAUCCUAAUCGUGGUGCUAUGCGUGUCGUUUAGCAGG CACUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUG GGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCGUGG UCUUUGAAUAAAGUCUGAGUGGGCGGC Chik- UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGG 391 Strain37997-E1 GAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCA (CHIKV E1 UGCUAUGGAACGAACAGCAGCCCCUGUUCUGGUUGCAGGCUCUUAUC antigen-Strain CCGCUGGCCGCCUUGAUCGUCCUGUGCAACUGUCUGAAACUCUUGCC 37997): AUGCUGCUGUAAGACCCUGGCUUUUUUAGCCGUAAUGAGCAUCGGUG CCCACACUGUGAGCGCGUACGAACACGUAACAGUGAUCCCGAACACG GUGGGAGUACCGUAUAAGACUCUUGUCAACAGACCGGGUUACAGCCC CAUGGUGUUGGAGAUGGAGCUACAAUCAGUCACCUUGGAACCAACAC UGUCACUUGACUACAUCACGUGCGAGUACAAAACUGUCAUCCCCUCC CCGUACGUGAAGUGCUGUGGUACAGCAGAGUGCAAGGACAAGAGCCU ACCAGACUACAGCUGCAAGGUCUUUACUGGAGUCUACCCAUUUAUGU GGGGCGGCGCCUACUGCUUUUGCGACGCCGAAAAUACGCAAUUGAGC GAGGCACAUGUAGAGAAAUCUGAAUCUUGCAAAACAGAGUUUGCAUC GGCCUACAGAGCCCACACCGCAUCGGCGUCGGCGAAGCUCCGCGUCCU UUACCAAGGAAACAACAUUACCGUAGCUGCCUACGCUAACGGUGACC AUGCCGUCACAGUAAAGGACGCCAAGUUUGUCGUGGGCCCAAUGUCC UCCGCCUGGACACCUUUUGACAACAAAAUCGUGGUGUACAAAGGCGA CGUCUACAACAUGGACUACCCACCUUUUGGCGCAGGAAGACCAGGAC AAUUUGGUGACAUUCAAAGUCGUACACCGGAAAGUAAAGACGUUUAU GCCAACACUCAGUUGGUACUACAGAGGCCAGCAGCAGGCACGGUACA UGUACCAUACUCUCAGGCACCAUCUGGCUUCAAGUAUUGGCUGAAGG AACGAGGAGCAUCGCUACAGCACACGGCACCGUUCGGUUGCCAGAUU GCGACAAACCCGGUAAGAGCUGUAAAUUGCGCUGUGGGGAACAUACC AAUUUCCAUCGACAUACCGGAUGCGGCCUUUACUAGGGUUGUCGAUG CACCCUCUGUAACGGACAUGUCAUGCGAAGUACCAGCCUGCACUCAC UCCUCCGACUUUGGGGGCGUCGCCAUCAUCAAAUACACAGCUAGCAA GAAAGGUAAAUGUGCAGUACAUUCGAUGACCAACGCCGUUACCAUUC GAGAAGCCGACGUAGAAGUAGAGGGGAACUCCCAGCUGCAAAUAUCC UUCUCAACAGCCCUGGCAAGCGCCGAGUUUCGCGUGCAAGUGUGCUC CACACAAGUACACUGCGCAGCCGCAUGCCACCCUCCAAAGGACCACAU AGUCAAUUACCCAGCAUCACACACCACCCUUGGGGUCCAGGAUAUAU CCACAACGGCAAUGUCUUGGGUGCAGAAGAUUACGGGAGGAGUAGGA UUAAUUGUUGCUGUUGCUGCCUUAAUUUUAAUUGUGGUGCUAUGCG UGUCGUUUAGCAGGCACUAAUGAUAAUAGGCUGGAGCCUCGGUGGCC AUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUG CACCCGUACCCCCGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC chikv-Brazillian- UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGG 392 E1 (CHIKV E1 GAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCA antigen- UGUACGAACACGUAACAGUGAUCCCGAACACGGUGGGAGUACCGUAU Brazilian strain) AAGACUCUAGUCAAUAGACCGGGCUACAGUCCCAUGGUAUUGGAGAU GGAACUACUGUCAGUCACUUUGGAGCCAACGCUAUCGCUUGAUUACA UCACGUGCGAGUACAAAACCGUUAUCCCGUCUCCGUACGUGAAAUGC UGCGGUACAGCAGAGUGCAAGGACAAAAACCUACCUGACUACAGCUG UAAGGUCUUCACCGGCGUCUACCCAUUUAUGUGGGGCGGAGCCUACU GCUUCUGCGACGCUGAAAACACGCAAUUGAGCGAAGCACACGUGGAG AAGUCCGAAUCAUGCAAAACAGAAUUUGCAUCAGCAUACAGGGCUCA UACCGCAUCCGCAUCAGCUAAGCUCCGCGUCCUUUACCAAGGAAAUA ACAUCACUGUAACUGCCUAUGCUAACGGCGACCAUGCCGUCACAGUU AAGGACGCCAAAUUCAUUGUGGGGCCAAUGUCUUCAGCCUGGACACC UUUCGACAACAAAAUUGUGGUGUACAAAGGUGACGUCUAUAACAUGG ACUACCCGCCCUUUGGCGCAGGAAGACCAGGACAAUUUGGCGAUAUC CAAAGUCGCACACCUGAGAGUAAAGACGUCUAUGCUAAUACACAACU GGUACUGCAGAGACCGGCUGCGGGUACGGUACAUGUGCCAUACUCUC AGGCACCAUCUGGCUUUAAGUAUUGGCUAAAAGAACGAGGGGCGUCG CUGCAGCACACAGCACCAUUUGGCUGCCAAAUAGCAACAAACCCGGU AAGAGCGGUGAAUUGCGCCGUAGGGAACAUGCCCAUCUCCAUCGACA UACCGGAUGCGGCCUUCAUUAGGGUCGUCGACGCGCCCUCUUUAACG GACAUGUCGUGCGAGGUACCAGCCUGCACCCAUUCCUCAGAUUUCGG GGGCGUCGCCAUUAUUAAAUAUGCAGCCAGCAAGAAAGGCAAGUGUG CGGUGCAUUCGAUGACCAACGCCGUCACAAUUCGGGAAGCUGAGAUA GAAGUUGAAGGGAAUUCUCAGCUGCAAAUCUCUUUCUCGACGGCCUU GGCCAGCGCCGAAUUCCGCGUACAAGUCUGUUCUACACAAGUACACU GUGUAGCCGAGUGCCACCCUCCGAAGGACCACAUAGUCAAUUACCCG GCGUCACAUACCACCCUCGGGGUCCAGGACAUUUCCGCUACGGCGCU GUCAUGGGUGCAGAAGAUCACGGGAGGCGUGGGACUGGUUGUCGCUG UUGCAGCACUGAUUCUAAUCGUGGUGCUAUGCGUGUCGUUCAGCAGG CACUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUG GGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCGUGG UCUUUGAAUAAAGUCUGAGUGGGCGGC ChiK.secE2 AUGGAGACCCCAGCUCAGCUUCUGUUUCUUCUCCUUCUAUGGCUGCC 393 HS3UPCRfree UGACACGACUGGACAUCACCACCAUCAUCAUAGUACAAAAGACAAUU (CHIKV secreted UCAAUGUGUACAAGGCCACCCGCCCUUAUUUAGCACACUGUCCAGAU E2 antigen): UGCGGUGAGGGGCACUCCUGUCACUCUCCUAUCGCCUUGGAGCGGAU CCGGAAUGAGGCGACCGAUGGAACACUGAAAAUCCAGGUAAGCUUGC AGAUUGGCAUCAAGACUGACGAUAGCCAUGAUUGGACCAAACUACGG UAUAUGGAUAGCCAUACACCUGCCGAUGCUGAACGGGCCGGUCUGCU UGUGAGAACUAGCGCUCCAUGCACCAUCACGGGGACAAUGGGACAUU UUAUCCUGGCUAGAUGCCCAAAGGGCGAAACCCUCACCGUCGGAUUC ACCGACUCAAGGAAAAUUUCUCACACAUGUACCCAUCCCUUCCACCA UGAGCCACCGGUGAUCGGGCGCGAACGCUUCCACAGCAGGCCUCAGC AUGGAAAAGAACUGCCAUGCUCGACCUAUGUACAGUCCACCGCCGCU ACCGCCGAAGAGAUCGAAGUGCAUAUGCCUCCCGACACACCCGACCG AACCCUAAUGACACAACAAUCUGGGAAUGUGAAGAUUACAGUCAAUG GACAGACUGUGAGGUAUAAGUGUAACUGCGGUGGCUCAAAUGAGGGC CUCACCACAACGGAUAAGGUGAUCAAUAACUGCAAAAUUGACCAGUG UCACGCGGCCGUGACCAACCAUAAGAACUGGCAGUACAACUCACCCU UAGUGCCUAGGAACGCUGAGCUGGGAGAUCGCAAGGGGAAGAUACAC AUUCCCUUCCCGUUGGCGAAUGUGACCUGCCGUGUGCCAAAAGCGAG AAAUCCUACCGUAACAUAUGGCAAAAAUCAGGUGACCAUGUUGCUCU ACCCGGAUCACCCAACUCUGCUGAGCUAUCGGAAUAUGGGACAAGAA CCCAAUUACCACGAGGAAUGGGUUACGCACAAGAAAGAGGUGACCCU UACAGUCCCUACUGAAGGUCUGGAAGUGACCUGGGGCAAUAACGAGC CUUAUAAGUACUGGCCCCAGAUGAGUACAAACGGCACCGCCCAUGGA CAUCCACACGAGAUCAUUCUGUAUUACUACGAACUAUAUCCCACAAU GACUGGCAAGCCUAUACCAAACCCACUUCUCGGCCUUGAUAGCACAU GAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCC UCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCGUGGUCUU UGAAUAAAGUCUGAGUGGGCGGC chikv-Brazillian- UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGG 394 E2 (CHIKV E2 GAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCA antigen- UGAGUACCAAGGACAACUUCAAUGUCUAUAAAGCCACAAGACCGUAC Brazilian strain): UUAGCUCACUGUCCCGACUGUGGAGAAGGGCACUCGUGCCAUAGUCC CGUAGCAUUAGAACGCAUCAGAAAUGAAGCGACAGACGGGACGCUGA AAAUCCAGGUCUCCUUGCAAAUCGGAAUAAAGACGGAUGAUAGCCAC GAUUGGACCAAGCUGCGUUACAUGGACAACCACACGCCAGCGGACGC AGAGAGGGCGGGGCUAUUUGUAAGAACAUCAGCACCGUGCACGAUUA CUGGAACAAUGGGACACUUCAUCCUGACCCGAUGUCCGAAAGGGGAA ACUCUGACGGUGGGAUUCACUGACAGUAGGAAGAUCAGUCACUCAUG UACGCACCCAUUUCACCACGACCCUCCUGUGAUAGGCCGGGAGAAAU UCCAUUCCCGACCGCAGCACGGUAAAGAGCUGCCUUGCAGCACGUAC GUGCAGAGCACCGCCGCAACUACCGAGGAGAUAGAGGUACACAUGCC CCCAGACACCCCUGAUCGCACAUUGAUGUCACAACAGUCCGGCAACG UAAAGAUCACAGUUAAUGGCCAGACGGUGCGGUACAAGUGUAAUUGC GGUGGCUCAAAUGAAGGACUAAUAACUACAGACAAAGUGAUUAAUA ACUGCAAAGUUGAUCAAUGUCAUGCCGCGGUCACCAAUCACAAAAAG UGGCAGUACAACUCCCCUCUGGUCCCGCGUAAUGCUGAACUUGGGGA CCGAAAAGGAAAAAUCCACAUCCCGUUUCCGCUGGCAAAUGUAACAU GCAGGGUGCCUAAAGCAAGGAACCCCACCGUGACGUACGGGAAAAAC CAAGUCAUCAUGCUACUGUAUCCCGACCACCCAACACUCCUGUCCUAC CGGAACAUGGGAGAAGAACCAAACUACCAAGAAGAGUGGGUGACGCA UAAGAAGGAAGUCGUGCUAACCGUGCCGACUGAAGGGCUCGAGGUCA CGUGGGGUAACAACGAGCCGUAUAAGUAUUGGCCGCAGUUAUCUACA AACGGUACAGCCCAUGGCCACCCGCAUGAGAUAAUUCUGUAUUAUUA UGAGCUGUACCCUACUAUGACUGUAGUAGUUGUGUCAGUGGCCUCGU UCGUACUCCUGUCGAUGGUGGGUGUGGCAGUGGGGAUGUGCAUGUGU GCACGACGCAGAUGCAUCACACCGUACGAACUGACACCAGGAGCUAC CGUCCCUUUCCUGCUUAGCCUAAUAUGCUGCAUCAGAACAGCUAAAG CGUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGG GCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCGUGGU CUUUGAAUAAAGUCUGAGUGGGCGGC chikv-Brazillian- UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGG 395 E2 (CHIKV E2 GAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCA antigen- UGAGUAUUAAGGACCACUUCAAUGUCUAUAAAGCCACAAGACCGUAC Brazilian strain): CUAGCUCACUGUCCCGACUGUGGAGAAGGGCACUCGUGCCAUAGUCC CGUAGCGCUAGAACGCAUCAGAAACGAAGCGACAGACGGGACGUUGA AAAUCCAGGUUUCCUUGCAAAUCGGAAUAAAGACGGAUGAUAGCCAU GAUUGGACCAAGCUGCGUUAUAUGGACAAUCACAUGCCAGCAGACGC AGAGCGGGCCGGGCUAUUUGUAAGAACGUCAGCACCGUGCACGAUUA CUGGAACAAUGGGACACUUCAUUCUGGCCCGAUGUCCGAAAGGAGAA ACUCUGACGGUGGGGUUCACUGACGGUAGGAAGAUCAGUCACUCAUG UACGCACCCAUUUCACCAUGACCCUCCUGUGAUAGGCCGGGAAAAAU UCCAUUCCCGACCGCAGCACGGUAGGGAACUACCUUGCAGCACGUAC GCGCAGAGCACCGCUGCAACUGCCGAGGAGAUAGAGGUACACAUGCC CCCAGACACCCCAGAUCGCACAUUAAUGUCACAACAGUCCGGCAAUG UAAAGAUCACAGUCAAUAGUCAGACGGUGCGGUACAAGUGCAAUUGU GGUGACUCAAGUGAAGGAUUAACCACUACAGAUAAAGUGAUUAAUA ACUGCAAGGUCGAUCAAUGCCAUGCCGCGGUCACCAAUCACAAAAAA UGGCAGUAUAACUCCCCUCUGGUCCCGCGUAAUGCUGAAUUCGGGGA CCGGAAAGGAAAAGUUCACAUUCCAUUUCCUCUGGCAAAUGUGACAU GCAGGGUGCCUAAAGCAAGAAACCCCACCGUGACGUACGGAAAAAAC CAAGUCAUCAUGUUGCUGUAUCCUGACCACCCAACGCUCCUGUCCUA CAGGAAUAUGGGAGAAGAACCAAACUAUCAAGAAGAGUGGGUGACGC AUAAGAAGGAGAUCAGGUUAACCGUGCCGACUGAGGGGCUCGAGGUC ACGUGGGGUAACAAUGAGCCGUACAAGUAUUGGCCGCAGUUAUCCAC AAACGGUACAGCCCACGGCCACCCGCAUGAGAUAAUUCUGUAUUAUU AUGAGCUGUACCCAACUAUGACUGCGGUAGUUUUGUCAGUGGCCUCG UUCAUACUCCUGUCGAUGGUGGGUGUGGCAGUGGGGAUGUGCAUGUG UGCACGACGCAGAUGCAUUACACCGUACGAACUGACACCAGGAGCUA CCGUCCCUUUCCUGCUUAGCCUAAUAUGCUGCAUUAGAACAGCUAAA GCGUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUG GGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCGUGG UCUUUGAAUAAAGUCUGAGUGGGCGGC Chik-Strain UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGG 396 37997-E2 GAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCA (CHIKV E2 UGCCAUAUCUAGCUCAUUGUCCUGACUGCGGAGAAGGGCAUUCGUGC Antigen-Strain CACAGCCCUAUCGCAUUGGAGCGCAUCAGAAAUGAAGCAACGGACGG 37997) AACGCUGAAAAUCCAGGUCUCUUUGCAGAUCGGGAUAAAGACAGAUG ACAGCCACGAUUGGACCAAGCUGCGCUAUAUGGAUAGCCAUACGCCA GCGGACGCGGAGCGAGCCGGAUUGCUUGUAAGGACUUCAGCACCGUG CACGAUCACCGGGACCAUGGGACACUUUAUUCUCGCCCGAUGCCCGA AAGGAGAGACGCUGACAGUGGGAUUUACGGACAGCAGAAAGAUCAGC CACACAUGCACACACCCGUUCCAUCAUGAACCACCUGUGAUAGGUAG GGAGAGGUUCCACUCUCGACCACAACAUGGUAAAGAGUUACCUUGCA GCACGUACGUGCAGAGCACCGCUGCCACUGCUGAGGAGAUAGAGGUG CAUAUGCCCCCAGAUACUCCUGACCGCACGCUGAUGACGCAGCAGUC UGGCAACGUGAAGAUCACAGUUAAUGGGCAGACGGUGCGGUACAAGU GCAACUGCGGUGGCUCAAACGAGGGACUGACAACCACAGACAAAGUG AUCAAUAACUGCAAAAUUGAUCAGUGCCAUGCUGCAGUCACUAAUCA CAAGAAUUGGCAAUACAACUCCCCUUUAGUCCCGCGCAACGCUGAAC UCGGGGACCGUAAAGGAAAGAUCCACAUCCCAUUCCCAUUGGCAAAC GUGACUUGCAGAGUGCCAAAAGCAAGAAACCCUACAGUAACUUACGG AAAAAACCAAGUCACCAUGCUGCUGUAUCCUGACCAUCCGACACUCU UGUCUUACCGUAACAUGGGACAGGAACCAAAUUACCACGAGGAGUGG GUGACACACAAGAAGGAGGUUACCUUGACCGUGCCUACUGAGGGUCU GGAGGUCACUUGGGGCAACAACGAACCAUACAAGUACUGGCCGCAGA UGUCUACGAACGGUACUGCUCAUGGUCACCCACAUGAGAUAAUCUUG UACUAUUAUGAGCUGUACCCCACUAUGACUGUAGUCAUUGUGUCGGU GGCCUCGUUCGUGCUUCUGUCGAUGGUGGGCACAGCAGUGGGAAUGU GUGUGUGCGCACGGCGCAGAUGCAUUACACCAUAUGAAUUAACACCA GGAGCCACUGUUCCCUUCCUGCUCAGCCUGCUAUGCUGCUGAUAAUA GGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCA GCCCCUCCUCCCCUUCCUGCACCCGUACCCCCGUGGUCUUUGAAUAAA GUCUGAGUGGGCGGC chikv-Brazillian- UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGG 397 E2-E1 (CHIKV GAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCA E1-E2 Antigen- UGAGUACCAAGGACAACUUCAAUGUCUAUAAAGCCACAAGACCGUAC Brazilian strain): UUAGCUCACUGUCCCGACUGUGGAGAAGGGCACUCGUGCCAUAGUCC CGUAGCAUUAGAACGCAUCAGAAAUGAAGCGACAGACGGGACGCUGA AAAUCCAGGUCUCCUUGCAAAUCGGAAUAAAGACGGAUGAUAGCCAC GAUUGGACCAAGCUGCGUUACAUGGACAACCACACGCCAGCGGACGC AGAGAGGGCGGGGCUAUUUGUAAGAACAUCAGCACCGUGCACGAUUA CUGGAACAAUGGGACACUUCAUCCUGACCCGAUGUCCGAAAGGGGAA ACUCUGACGGUGGGAUUCACUGACAGUAGGAAGAUCAGUCACUCAUG UACGCACCCAUUUCACCACGACCCUCCUGUGAUAGGCCGGGAGAAAU UCCAUUCCCGACCGCAGCACGGUAAAGAGCUGCCUUGCAGCACGUAC GUGCAGAGCACCGCCGCAACUACCGAGGAGAUAGAGGUACACAUGCC CCCAGACACCCCUGAUCGCACAUUGAUGUCACAACAGUCCGGCAACG UAAAGAUCACAGUUAAUGGCCAGACGGUGCGGUACAAGUGUAAUUGC GGUGGCUCAAAUGAAGGACUAAUAACUACAGACAAAGUGAUUAAUA ACUGCAAAGUUGAUCAAUGUCAUGCCGCGGUCACCAAUCACAAAAAG UGGCAGUACAACUCCCCUCUGGUCCCGCGUAAUGCUGAACUUGGGGA CCGAAAAGGAAAAAUCCACAUCCCGUUUCCGCUGGCAAAUGUAACAU GCAGGGUGCCUAAAGCAAGGAACCCCACCGUGACGUACGGGAAAAAC CAAGUCAUCAUGCUACUGUAUCCCGACCACCCAACACUCCUGUCCUAC CGGAACAUGGGAGAAGAACCAAACUACCAAGAAGAGUGGGUGACGCA UAAGAAGGAAGUCGUGCUAACCGUGCCGACUGAAGGGCUCGAGGUCA CGUGGGGUAACAACGAGCCGUAUAAGUAUUGGCCGCAGUUAUCUACA AACGGUACAGCCCAUGGCCACCCGCAUGAGAUAAUUCUGUAUUAUUA UGAGCUGUACCCUACUAUGACUGUAGUAGUUGUGUCAGUGGCCUCGU UCGUACUCCUGUCGAUGGUGGGUGUGGCAGUGGGGAUGUGCAUGUGU GCACGACGCAGAUGCAUCACACCGUACGAACUGACACCAGGAGCUAC CGUCCCUUUCCUGCUUAGCCUAAUAUGCUGCAUCAGAACAGCUAAAG CGUACGAACACGUAACAGUGAUCCCGAACACGGUGGGAGUACCGUAU AAGACUCUAGUCAAUAGACCGGGCUACAGUCCCAUGGUAUUGGAGAU GGAACUACUGUCAGUCACUUUGGAGCCAACGCUAUCGCUUGAUUACA UCACGUGCGAGUACAAAACCGUUAUCCCGUCUCCGUACGUGAAAUGC UGCGGUACAGCAGAGUGCAAGGACAAAAACCUACCUGACUACAGCUG UAAGGUCUUCACCGGCGUCUACCCAUUUAUGUGGGGCGGAGCCUACU GCUUCUGCGACGCUGAAAACACGCAAUUGAGCGAAGCACACGUGGAG AAGUCCGAAUCAUGCAAAACAGAAUUUGCAUCAGCAUACAGGGCUCA UACCGCAUCCGCAUCAGCUAAGCUCCGCGUCCUUUACCAAGGAAAUA ACAUCACUGUAACUGCCUAUGCUAACGGCGACCAUGCCGUCACAGUU AAGGACGCCAAAUUCAUUGUGGGGCCAAUGUCUUCAGCCUGGACACC UUUCGACAACAAAAUUGUGGUGUACAAAGGUGACGUCUAUAACAUGG ACUACCCGCCCUUUGGCGCAGGAAGACCAGGACAAUUUGGCGAUAUC CAAAGUCGCACACCUGAGAGUAAAGACGUCUAUGCUAAUACACAACU GGUACUGCAGAGACCGGCUGCGGGUACGGUACAUGUGCCAUACUCUC AGGCACCAUCUGGCUUUAAGUAUUGGCUAAAAGAACGAGGGGCGUCG CUGCAGCACACAGCACCAUUUGGCUGCCAAAUAGCAACAAACCCGGU AAGAGCGGUGAAUUGCGCCGUAGGGAACAUGCCCAUCUCCAUCGACA UACCGGAUGCGGCCUUCAUUAGGGUCGUCGACGCGCCCUCUUUAACG GACAUGUCGUGCGAGGUACCAGCCUGCACCCAUUCCUCAGAUUUCGG GGGCGUCGCCAUUAUUAAAUAUGCAGCCAGCAAGAAAGGCAAGUGUG CGGUGCAUUCGAUGACCAACGCCGUCACAAUUCGGGAAGCUGAGAUA GAAGUUGAAGGGAAUUCUCAGCUGCAAAUCUCUUUCUCGACGGCCUU GGCCAGCGCCGAAUUCCGCGUACAAGUCUGUUCUACACAAGUACACU GUGUAGCCGAGUGCCACCCUCCGAAGGACCACAUAGUCAAUUACCCG GCGUCACAUACCACCCUCGGGGUCCAGGACAUUUCCGCUACGGCGCU GUCAUGGGUGCAGAAGAUCACGGGAGGCGUGGGACUGGUUGUCGCUG UUGCAGCACUGAUUCUAAUCGUGGUGCUAUGCGUGUCGUUCAGCAGG CACUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCCCUUG GGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCCGUGG UCUUUGAAUAAAGUCUGAGUGGGCGGC chikv-Brazillian- UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGG 398 E2-E1 (CHIKV GAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCA E1-E2 Antigen- UGAGUAUUAAGGACCACUUCAAUGUCUAUAAAGCCACAAGACCGUAC Brazilian strain): CUAGCUCACUGUCCCGACUGUGGAGAAGGGCACUCGUGCCAUAGUCC CGUAGCGCUAGAACGCAUCAGAAACGAAGCGACAGACGGGACGUUGA AAAUCCAGGUUUCCUUGCAAAUCGGAAUAAAGACGGAUGAUAGCCAU GAUUGGACCAAGCUGCGUUAUAUGGACAAUCACAUGCCAGCAGACGC AGAGCGGGCCGGGCUAUUUGUAAGAACGUCAGCACCGUGCACGAUUA CUGGAACAAUGGGACACUUCAUUCUGGCCCGAUGUCCGAAAGGAGAA ACUCUGACGGUGGGGUUCACUGACGGUAGGAAGAUCAGUCACUCAUG UACGCACCCAUUUCACCAUGACCCUCCUGUGAUAGGCCGGGAAAAAU UCCAUUCCCGACCGCAGCACGGUAGGGAACUACCUUGCAGCACGUAC GCGCAGAGCACCGCUGCAACUGCCGAGGAGAUAGAGGUACACAUGCC CCCAGACACCCCAGAUCGCACAUUAAUGUCACAACAGUCCGGCAAUG UAAAGAUCACAGUCAAUAGUCAGACGGUGCGGUACAAGUGCAAUUGU GGUGACUCAAGUGAAGGAUUAACCACUACAGAUAAAGUGAUUAAUA ACUGCAAGGUCGAUCAAUGCCAUGCCGCGGUCACCAAUCACAAAAAA UGGCAGUAUAACUCCCCUCUGGUCCCGCGUAAUGCUGAAUUCGGGGA CCGGAAAGGAAAAGUUCACAUUCCAUUUCCUCUGGCAAAUGUGACAU GCAGGGUGCCUAAAGCAAGAAACCCCACCGUGACGUACGGAAAAAAC CAAGUCAUCAUGUUGCUGUAUCCUGACCACCCAACGCUCCUGUCCUA CAGGAAUAUGGGAGAAGAACCAAACUAUCAAGAAGAGUGGGUGACGC AUAAGAAGGAGAUCAGGUUAACCGUGCCGACUGAGGGGCUCGAGGUC ACGUGGGGUAACAAUGAGCCGUACAAGUAUUGGCCGCAGUUAUCCAC AAACGGUACAGCCCACGGCCACCCGCAUGAGAUAAUUCUGUAUUAUU AUGAGCUGUACCCAACUAUGACUGCGGUAGUUUUGUCAGUGGCCUCG UUCAUACUCCUGUCGAUGGUGGGUGUGGCAGUGGGGAUGUGCAUGUG UGCACGACGCAGAUGCAUUACACCGUACGAACUGACACCAGGAGCUA CCGUCCCUUUCCUGCUUAGCCUAAUAUGCUGCAUUAGAACAGCUAAA GCGUACGAACACGUAACAGUGAUCCCGAACACGGUGGGAGUACCGUA UAAGACUCUAGUCAACAGACCGGGCUACAGCCCCAUGGUAUUGGAGA UGGAGCUUCUGUCUGUCACCUUGGAACCAACGCUAUCGCUUGAUUAC AUCACGUGCGAGUAUAAAACCGUUAUCCCGUCUCCGUACGUGAAAUG CUGCGGUACAGCAGAGUGUAAGGACAAGAGCCUACCUGAUUACAGCU GUAAGGUCUUCACCGGCGUCUACCCAUUCAUGUGGGGCGGCGCCUAC UGCUUCUGCGACACCGAAAAUACGCAAUUGAGCGAAGCACAUGUGGA GAAGUCCGAAUCAUGCAAAACAGAAUUUGCAUCAGCAUACAGGGCUC AUACCGCAUCCGCAUCAGCUAAGCUCCGCGUCCUUUACCAAGGAAAU AAUAUCACUGUGGCUGCUUAUGCAAACGGCGACCAUGCCGUCACAGU UAAGGACGCUAAAUUCAUAGUGGGGCCAAUGUCUUCAGCCUGGACAC CUUUCGACAAUAAAAUCGUGGUGUACAAAGGCGACGUCUACAACAUG GACUACCCGCCCUUCGGCGCAGGAAGACCAGGACAAUUUGGCGACAU CCAAAGUCGCACGCCUGAGAGCGAAGACGUCUAUGCUAAUACACAAC UGGUACUGCAGAGACCGUCCGCGGGUACGGUGCACGUGCCGUACUCU CAGGCACCAUCUGGCUUCAAGUAUUGGCUAAAAGAACGAGGGGCGUC GCUGCAGCACACAGCACCAUUUGGCUGUCAAAUAGCAACAAACCCGG UAAGAGCGAUGAACUGCGCCGUAGGGAACAUGCCUAUCUCCAUCGAC AUACCGGACGCGGCCUUUACCAGGGUCGUCGACGCGCCAUCUUUAAC GGACAUGUCGUGUGAGGUAUCAGCCUGCACCCAUUCCUCAGACUUUG GGGGCGUAGCCAUCAUUAAAUAUGCAGCCAGUAAGAAAGGCAAGUGU GCAGUGCACUCGAUGACUAACGCCGUCACUAUUCGGGAAGCUGAAAU AGAAGUAGAAGGGAACUCUCAGUUGCAAAUCUCUUUUUCGACGGCCC UAGCCAGCGCCGAAUUUCGCGUACAAGUCUGUUCUACACAAGUACAC UGUGCAGCCGAGUGCCAUCCACCGAAAGACCAUAUAGUCAAUUACCC GGCGUCACACACCACCCUCGGGGUCCAAGACAUUUCCGCUACGGCGA UGUCAUGGGUGCAGAAGAUCACGGGAGGUGUGGGACUGGUUGUCGCU GUUGCAGCACUGAUCCUAAUCGUGGUGCUAUGCGUGUCGUUUAGCAG GCACAUGAGUAUUAAGGACCACUUCAAUGUCUAUAAAGCCACAAGAC CGUACCUAGCUCACUGUCCCGACUGUGGAGAAGGGCACUCGUGCCAU AGUCCCGUAGCGCUAGAACGCAUCAGAAACGAAGCGACAGACGGGAC GUUGAAAAUCCAGGUUUCCUUGCAAAUCGGAAUAAAGACGGAUGAUA GCCAUGAUUGGACCAAGCUGCGUUAUAUGGACAAUCACAUGCCAGCA GACGCAGAGCGGGCCGGGCUAUUUGUAAGAACGUCAGCACCGUGCAC GAUUACUGGAACAAUGGGACACUUCAUUCUGGCCCGAUGUCCGAAAG GAGAAACUCUGACGGUGGGGUUCACUGACGGUAGGAAGAUCAGUCAC UCAUGUACGCACCCAUUUCACCAUGACCCUCCUGUGAUAGGCCGGGA AAAAUUCCAUUCCCGACCGCAGCACGGUAGGGAACUACCUUGCAGCA CGUACGCGCAGAGCACCGCUGCAACUGCCGAGGAGAUAGAGGUACAC AUGCCCCCAGACACCCCAGAUCGCACAUUAAUGUCACAACAGUCCGG CAAUGUAAAGAUCACAGUCAAUAGUCAGACGGUGCGGUACAAGUGCA AUUGUGGUGACUCAAGUGAAGGAUUAACCACUACAGAUAAAGUGAU UAAUAACUGCAAGGUCGAUCAAUGCCAUGCCGCGGUCACCAAUCACA AAAAAUGGCAGUAUAACUCCCCUCUGGUCCCGCGUAAUGCUGAAUUC GGGGACCGGAAAGGAAAAGUUCACAUUCCAUUUCCUCUGGCAAAUGU GACAUGCAGGGUGCCUAAAGCAAGAAACCCCACCGUGACGUACGGAA AAAACCAAGUCAUCAUGUUGCUGUAUCCUGACCACCCAACGCUCCUG UCCUACAGGAAUAUGGGAGAAGAACCAAACUAUCAAGAAGAGUGGGU GACGCAUAAGAAGGAGAUCAGGUUAACCGUGCCGACUGAGGGGCUCG AGGUCACGUGGGGUAACAAUGAGCCGUACAAGUAUUGGCCGCAGUUA UCCACAAACGGUACAGCCCACGGCCACCCGCAUGAGAUAAUUCUGUA UUAUUAUGAGCUGUACCCAACUAUGACUGCGGUAGUUUUGUCAGUGG CCUCGUUCAUACUCCUGUCGAUGGUGGGUGUGGCAGUGGGGAUGUGC AUGUGUGCACGACGCAGAUGCAUUACACCGUACGAACUGACACCAGG AGCUACCGUCCCUUUCCUGCUUAGCCUAAUAUGCUGCAUUAGAACAG CUAAAGCGUACGAACACGUAACAGUGAUCCCGAACACGGUGGGAGUA CCGUAUAAGACUCUAGUCAACAGACCGGGCUACAGCCCCAUGGUAUU GGAGAUGGAGCUUCUGUCUGUCACCUUGGAACCAACGCUAUCGCUUG AUUACAUCACGUGCGAGUAUAAAACCGUUAUCCCGUCUCCGUACGUG AAAUGCUGCGGUACAGCAGAGUGUAAGGACAAGAGCCUACCUGAUUA CAGCUGUAAGGUCUUCACCGGCGUCUACCCAUUCAUGUGGGGCGGCG CCUACUGCUUCUGCGACACCGAAAAUACGCAAUUGAGCGAAGCACAU GUGGAGAAGUCCGAAUCAUGCAAAACAGAAUUUGCAUCAGCAUACAG GGCUCAUACCGCAUCCGCAUCAGCUAAGCUCCGCGUCCUUUACCAAG GAAAUAAUAUCACUGUGGCUGCUUAUGCAAACGGCGACCAUGCCGUC ACAGUUAAGGACGCUAAAUUCAUAGUGGGGCCAAUGUCUUCAGCCUG GACACCUUUCGACAAUAAAAUCGUGGUGUACAAAGGCGACGUCUACA ACAUGGACUACCCGCCCUUCGGCGCAGGAAGACCAGGACAAUUUGGC GACAUCCAAAGUCGCACGCCUGAGAGCGAAGACGUCUAUGCUAAUAC ACAACUGGUACUGCAGAGACCGUCCGCGGGUACGGUGCACGUGCCGU ACUCUCAGGCACCAUCUGGCUUCAAGUAUUGGCUAAAAGAACGAGGG GCGUCGCUGCAGCACACAGCACCAUUUGGCUGUCAAAUAGCAACAAA CCCGGUAAGAGCGAUGAACUGCGCCGUAGGGAACAUGCCUAUCUCCA UCGACAUACCGGACGCGGCCUUUACCAGGGUCGUCGACGCGCCAUCU UUAACGGACAUGUCGUGUGAGGUAUCAGCCUGCACCCAUUCCUCAGA CUUUGGGGGCGUAGCCAUCAUUAAAUAUGCAGCCAGUAAGAAAGGCA AGUGUGCAGUGCACUCGAUGACUAACGCCGUCACUAUUCGGGAAGCU GAAAUAGAAGUAGAAGGGAACUCUCAGUUGCAAAUCUCUUUUUCGAC GGCCCUAGCCAGCGCCGAAUUUCGCGUACAAGUCUGUUCUACACAAG UACACUGUGCAGCCGAGUGCCAUCCACCGAAAGACCAUAUAGUCAAU UACCCGGCGUCACACACCACCCUCGGGGUCCAAGACAUUUCCGCUACG GCGAUGUCAUGGGUGCAGAAGAUCACGGGAGGUGUGGGACUGGUUG UCGCUGUUGCAGCACUGAUCCUAAUCGUGGUGCUAUGCGUGUCGUUU AGCAGGCACUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGC CCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCC CCGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC Chik-Strain UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGG 399 37997-E2-E1 GAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCA (CHIKV E1-E2 UGCCAUAUCUAGCUCAUUGUCCUGACUGCGGAGAAGGGCAUUCGUGC Antigen-Strain CACAGCCCUAUCGCAUUGGAGCGCAUCAGAAAUGAAGCAACGGACGG 37997): AACGCUGAAAAUCCAGGUCUCUUUGCAGAUCGGGAUAAAGACAGAUG ACAGCCACGAUUGGACCAAGCUGCGCUAUAUGGAUAGCCAUACGCCA GCGGACGCGGAGCGAGCCGGAUUGCUUGUAAGGACUUCAGCACCGUG CACGAUCACCGGGACCAUGGGACACUUUAUUCUCGCCCGAUGCCCGA AAGGAGAGACGCUGACAGUGGGAUUUACGGACAGCAGAAAGAUCAGC CACACAUGCACACACCCGUUCCAUCAUGAACCACCUGUGAUAGGUAG GGAGAGGUUCCACUCUCGACCACAACAUGGUAAAGAGUUACCUUGCA GCACGUACGUGCAGAGCACCGCUGCCACUGCUGAGGAGAUAGAGGUG CAUAUGCCCCCAGAUACUCCUGACCGCACGCUGAUGACGCAGCAGUC UGGCAACGUGAAGAUCACAGUUAAUGGGCAGACGGUGCGGUACAAGU GCAACUGCGGUGGCUCAAACGAGGGACUGACAACCACAGACAAAGUG AUCAAUAACUGCAAAAUUGAUCAGUGCCAUGCUGCAGUCACUAAUCA CAAGAAUUGGCAAUACAACUCCCCUUUAGUCCCGCGCAACGCUGAAC UCGGGGACCGUAAAGGAAAGAUCCACAUCCCAUUCCCAUUGGCAAAC GUGACUUGCAGAGUGCCAAAAGCAAGAAACCCUACAGUAACUUACGG AAAAAACCAAGUCACCAUGCUGCUGUAUCCUGACCAUCCGACACUCU UGUCUUACCGUAACAUGGGACAGGAACCAAAUUACCACGAGGAGUGG GUGACACACAAGAAGGAGGUUACCUUGACCGUGCCUACUGAGGGUCU GGAGGUCACUUGGGGCAACAACGAACCAUACAAGUACUGGCCGCAGA UGUCUACGAACGGUACUGCUCAUGGUCACCCACAUGAGAUAAUCUUG UACUAUUAUGAGCUGUACCCCACUAUGACUGUAGUCAUUGUGUCGGU GGCCUCGUUCGUGCUUCUGUCGAUGGUGGGCACAGCAGUGGGAAUGU GUGUGUGCGCACGGCGCAGAUGCAUUACACCAUAUGAAUUAACACCA GGAGCCACUGUUCCCUUCCUGCUCAGCCUGCUAUGCUGCCUAUGGAA CGAACAGCAGCCCCUGUUCUGGUUGCAGGCUCUUAUCCCGCUGGCCG CCUUGAUCGUCCUGUGCAACUGUCUGAAACUCUUGCCAUGCUGCUGU AAGACCCUGGCUUUUUUAGCCGUAAUGAGCAUCGGUGCCCACACUGU GAGCGCGUACGAACACGUAACAGUGAUCCCGAACACGGUGGGAGUAC CGUAUAAGACUCUUGUCAACAGACCGGGUUACAGCCCCAUGGUGUUG GAGAUGGAGCUACAAUCAGUCACCUUGGAACCAACACUGUCACUUGA CUACAUCACGUGCGAGUACAAAACUGUCAUCCCCUCCCCGUACGUGA AGUGCUGUGGUACAGCAGAGUGCAAGGACAAGAGCCUACCAGACUAC AGCUGCAAGGUCUUUACUGGAGUCUACCCAUUUAUGUGGGGCGGCGC CUACUGCUUUUGCGACGCCGAAAAUACGCAAUUGAGCGAGGCACAUG UAGAGAAAUCUGAAUCUUGCAAAACAGAGUUUGCAUCGGCCUACAGA GCCCACACCGCAUCGGCGUCGGCGAAGCUCCGCGUCCUUUACCAAGG AAACAACAUUACCGUAGCUGCCUACGCUAACGGUGACCAUGCCGUCA CAGUAAAGGACGCCAAGUUUGUCGUGGGCCCAAUGUCCUCCGCCUGG ACACCUUUUGACAACAAAAUCGUGGUGUACAAAGGCGACGUCUACAA CAUGGACUACCCACCUUUUGGCGCAGGAAGACCAGGACAAUUUGGUG ACAUUCAAAGUCGUACACCGGAAAGUAAAGACGUUUAUGCCAACACU CAGUUGGUACUACAGAGGCCAGCAGCAGGCACGGUACAUGUACCAUA CUCUCAGGCACCAUCUGGCUUCAAGUAUUGGCUGAAGGAACGAGGAG CAUCGCUACAGCACACGGCACCGUUCGGUUGCCAGAUUGCGACAAAC CCGGUAAGAGCUGUAAAUUGCGCUGUGGGGAACAUACCAAUUUCCAU CGACAUACCGGAUGCGGCCUUUACUAGGGUUGUCGAUGCACCCUCUG UAACGGACAUGUCAUGCGAAGUACCAGCCUGCACUCACUCCUCCGAC UUUGGGGGCGUCGCCAUCAUCAAAUACACAGCUAGCAAGAAAGGUAA AUGUGCAGUACAUUCGAUGACCAACGCCGUUACCAUUCGAGAAGCCG ACGUAGAAGUAGAGGGGAACUCCCAGCUGCAAAUAUCCUUCUCAACA GCCCUGGCAAGCGCCGAGUUUCGCGUGCAAGUGUGCUCCACACAAGU ACACUGCGCAGCCGCAUGCCACCCUCCAAAGGACCACAUAGUCAAUU ACCCAGCAUCACACACCACCCUUGGGGUCCAGGAUAUAUCCACAACG GCAAUGUCUUGGGUGCAGAAGAUUACGGGAGGAGUAGGAUUAAUUG UUGCUGUUGCUGCCUUAAUUUUAAUUGUGGUGCUAUGCGUGUCGUUU AGCAGGCACUAAUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCU UGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUA CCCCCGUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC Chik.C-E3-E2- UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGG 400 6K- GAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCA E1_HS3UPCRfree UGGAGUUUAUCCCUACGCAGACGUUCUAUAAUCGGAGGUACCAGCCC (C-E3-E2-6K- AGGCCUUGGGCCCCCCGCCCUACAAUCCAAGUGAUAAGACCACGUCCC E1 Antigen) AGGCCGCAGAGACAAGCCGGCCAAUUGGCGCAACUCAUCAGCGCAGU UAACAAGUUGACCAUGCGAGCGGUUCCUCAGCAGAAGCCGAGGCGGA ACCGGAAGAAUAAGAAACAACGCCAAAAGAAACAGGCGCCGCAGAAC GACCCUAAACAGAAGAAACAACCUCCCCAGAAAAAGCCAGCUCAGAA GAAGAAGAAGCCUGGACGCCGUGAAAGAAUGUGCAUGAAAAUCGAAA AUGAUUGCAUCUUUGAGGUGAAGCACGAGGGCAAAGUGAUGGGGUA CGCAUGCCUGGUGGGCGAUAAGGUCAUGAAGCCAGCACAUGUGAAGG GGACAAUCGAUAAUGCUGAUCUGGCCAAGCUAGCUUUUAAACGUAGC UCCAAAUACGAUCUUGAGUGUGCCCAGAUACCUGUGCACAUGAAAUC UGAUGCAAGCAAGUUCACACACGAGAAGCCUGAGGGCUAUUAUAACU GGCAUCAUGGUGCGGUUCAGUACUCCGGCGGCCGAUUUACCAUUCCU ACAGGGGCAGGAAAGCCGGGCGAUUCGGGGAGACCCAUUUUCGACAA CAAAGGCCGCGUGGUAGCUAUCGUGCUCGGUGGGGCUAAUGAGGGUG CACGUACUGCACUUAGCGUGGUUACCUGGAAUAAGGACAUUGUCACA AAGAUUACACCGGAGGGAGCAGAGGAAUGGAGCCUGGCACUGCCCGU UCUGUGCCUGCUGGCCAACACCACUUUCCCAUGUAGUCAACCCCCUU GCACUCCCUGCUGCUAUGAGAAAGAGCCUGAGAGCACGUUACGUAUG CUGGAAGAUAAUGUCAUGAGGCCCGGGUACUAUCAACUGCUCAAGGC UAGUCUGACAUGCUCGCCCCACAGGCAGCGCAGGUCCACGAAAGAUA ACUUCAACGUUUACAAGGCUACUAGGCCUUAUUUGGCCCACUGUCCC GAUUGCGGAGAGGGACAUUCUUGUCAUAGUCCUAUUGCCUUGGAGCG AAUCCGCAACGAGGCCACUGAUGGAACCCUUAAGAUUCAAGUAUCUU UGCAGAUUGGCAUUAAGACAGAUGAUUCCCAUGACUGGACAAAGCUU CGGUACAUGGACUCACACACGCCUGCAGAUGCUGAAAGGGCAGGGCU CUUGGUCAGGACCUCGGCCCCUUGUACAAUUACCGGGACCAUGGGCC ACUUCAUCCUUGCACGCUGCCCUAAGGGGGAGACGCUGACGGUGGGC UUUACUGACUCGCGUAAGAUCUCACACACAUGUACACACCCUUUCCA CCACGAACCUCCAGUCAUAGGGAGAGAGAGAUUUCACUCUCGCCCAC AGCAUGGCAAAGAGCUGCCAUGCUCCACAUAUGUCCAGAGCACUGCU GCUACCGCUGAAGAAAUUGAGGUUCACAUGCCACCCGAUACACCAGA CCGUACUCUGAUGACCCAACAGAGCGGCAACGUGAAGAUUACCGUAA AUGGACAGACCGUGAGAUAUAAGUGCAACUGUGGUGGCUCCAAUGAG GGCUUAACAACAACGGAUAAGGUGAUUAACAAUUGCAAAAUAGAUCA GUGCCAUGCCGCAGUGACCAAUCACAAGAAUUGGCAAUACAACUCAC CCCUAGUGCCGAGGAACGCAGAACUAGGCGACAGGAAAGGGAAAAUC CAUAUACCCUUCCCCCUAGCAAAUGUGACCUGCCGAGUGCCCAAGGC CAGAAACCCCACGGUUACUUACGGCAAGAACCAGGUGACGAUGCUUU UGUACCCAGACCAUCCCACCUUGCUCUCUUAUAGAAACAUGGGACAG GAGCCUAACUAUCAUGAGGAGUGGGUGACACACAAGAAAGAAGUCAC CCUUACCGUGCCUACCGAAGGGCUUGAAGUCACCUGGGGCAACAACG AGCCUUACAAGUAUUGGCCACAGAUGUCCACAAACGGAACAGCCCAC GGCCACCCGCACGAGAUCAUACUGUAUUACUAUGAGCUUUAUCCCAC AAUGACUGUCGUAAUUGUGAGCGUUGCCAGCUUCGUGUUGCUUUCAA UGGUUGGCACUGCCGUGGGGAUGUGCGUGUGUGCUAGGCGCCGCUGU AUAACUCCUUAUGAACUAACUCCAGGCGCCACCGUUCCUUUCCUGCU CUCACUACUGUGUUGUGUGCGCACAACAAAGGCUGCCACCUACUACG AAGCCGCCGCCUACUUAUGGAAUGAACAGCAGCCUCUCUUUUGGUUA CAGGCGCUGAUUCCUCUUGCUGCCCUGAUCGUGCUAUGCAACUGCCU CAAGCUGCUGCCCUGUUGUUGCAAGACCCUAGCUUUUCUCGCCGUGA UGAGCAUCGGGGCACAUACAGUGUCCGCCUAUGAGCACGUCACCGUU AUCCCGAACACCGUCGGUGUGCCAUAUAAGACGUUAGUCAAUCGACC CGGCUACUCUCCAAUGGUGCUGGAAAUGGAACUCCAGAGUGUGACAC UGGAGCCAACCUUAUCCCUCGAUUAUAUUACCUGCGAAUACAAGACC GUCAUCCCUUCACCCUAUGUCAAGUGCUGUGGGACCGCUGAAUGCAA AGACAAGAGCUUGCCUGAUUACAGUUGCAAGGUCUUCACAGGUGUGU ACCCCUUCAUGUGGGGGGGAGCUUAUUGCUUUUGUGAUGCUGAGAAC ACCCAACUGAGCGAGGCUCACGUCGAGAAAUCUGAGUCUUGCAAGAC CGAGUUUGCCUCAGCUUACAGGGCCCACACGGCCAGCGCAUCCGCCA AAUUGAGGGUACUCUACCAGGGUAAUAAUAUCACCGUUGCCGCAUAU GCAAACGGCGAUCACGCCGUGACUGUCAAGGAUGCCAAGUUCGUUGU GGGCCCCAUGUCUAGCGCUUGGACACCGUUCGAUAAUAAGAUCGUCG UGUACAAAGGGGACGUGUAUAAUAUGGACUACCCACCUUUCGGGGCC GGCCGACCGGGACAGUUCGGGGAUAUUCAGAGCCGCACACCCGAAUC UAAAGAUGUUUACGCCAAUACUCAGCUCGUCCUGCAGAGGCCCGCCG CUGGUACAGUUCACGUUCCUUACUCACAGGCACCCUCUGGGUUUAAG UAUUGGCUGAAAGAACGAGGUGCCAGCUUGCAGCAUACAGCGCCUUU CGGAUGCCAGAUUGCCACUAACCCCGUACGGGCUGUCAACUGCGCGG UCGGCAAUAUUCCCAUUAGCAUUGAUAUCCCGGACGCAGCUUUCACC AGGGUUGUGGACGCCCCGAGCGUCACCGACAUGAGUUGUGAGGUGCC AGCCUGCACGCAUAGCAGUGAUUUCGGCGGCGUCGCCAUCAUUAAAU AUACCGCAAGCAAGAAAGGCAAGUGCGCCGUCCACUCGAUGACUAAC GCCGUCACAAUUCGGGAAGCCGAUGUUGAGGUCGAAGGCAACUCCCA GCUGCAGAUCAGCUUCUCUACUGCUCUUGCAAGCGCCGAGUUUCGAG UCCAGGUCUGCAGUACGCAGGUGCAUUGUGCAGCUGCCUGCCAUCCA CCCAAAGAUCAUAUUGUGAAUUAUCCGGCGUCACAUACCACACUGGG GGUCCAGGAUAUUAGUACAACGGCGAUGUCCUGGGUGCAGAAAAUUA CGGGAGGAGUGGGCUUAAUUGUUGCCGUGGCGGCCCUGAUCCUGAUC GUUGUGCUGUGUGUUAGCUUCUCUAGGCAUGACUAUAAAGAUGACGA UGACAAAUGAUAAUAGGCUGGAGCCUCGGUGGCCAUGCUUCUUGCCC CUUGGGCCUCCCCCCAGCCCCUCCUCCCCUUCCUGCACCCGUACCCCC GUGGUCUUUGAAUAAAGUCUGAGUGGGCGGC CHIKV C-E3- UCAAGCUUUUGGACCCUCGUACAGAAGCUAAUACGACUCACUAUAGG 401 E2-6K-E1 GAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCCACCA UGGAGUUUAUCCCUACGCAGACGUUCUAUAAUCGGAGGUACCAGCCC AGGCCUUGGGCCCCCCGCCCUACAAUCCAAGUGAUAAGACCACGUCCC AGGCCGCAGAGACAAGCCGGCCAAUUGGCGCAACUCAUCAGCGCAGU UAACAAGUUGACCAUGCGAGCGGUUCCUCAGCAGAAGCCGAGGCGGA ACCGGAAGAAUAAGAAACAACGCCAAAAGAAACAGGCGCCGCAGAAC GACCCUAAACAGAAGAAACAACCUCCCCAGAAAAAGCCAGCUCAGAA GAAGAAGAAGCCUGGACGCCGUGAAAGAAUGUGCAUGAAAAUCGAAA AUGAUUGCAUCUUUGAGGUGAAGCACGAGGGCAAAGUGAUGGGGUA CGCAUGCCUGGUGGGCGAUAAGGUCAUGAAGCCAGCACAUGUGAAGG GGACAAUCGAUAAUGCUGAUCUGGCCAAGCUAGCUUUUAAACGUAGC UCCAAAUACGAUCUUGAGUGUGCCCAGAUACCUGUGCACAUGAAAUC UGAUGCAAGCAAGUUCACACACGAGAAGCCUGAGGGCUAUUAUAACU GGCAUCAUGGUGCGGUUCAGUACUCCGGCGGCCGAUUUACCAUUCCU ACAGGGGCAGGAAAGCCGGGCGAUUCGGGGAGACCCAUUUUCGACAA CAAAGGCCGCGUGGUAGCUAUCGUGCUCGGUGGGGCUAAUGAGGGUG CACGUACUGCACUUAGCGUGGUUACCUGGAAUAAGGACAUUGUCACA AAGAUUACACCGGAGGGAGCAGAGGAAUGGAGCCUGGCACUGCCCGU UCUGUGCCUGCUGGCCAACACCACUUUCCCAUGUAGUCAACCCCCUU GCACUCCCUGCUGCUAUGAGAAAGAGCCUGAGAGCACGUUACGUAUG CUGGAAGAUAAUGUCAUGAGGCCCGGGUACUAUCAACUGCUCAAGGC UAGUCUGACAUGCUCGCCCCACAGGCAGCGCAGGUCCACGAAAGAUA ACUUCAACGUUUACAAGGCUACUAGGCCUUAUUUGGCCCACUGUCCC GAUUGCGGAGAGGGACAUUCUUGUCAUAGUCCUAUUGCCUUGGAGCG AAUCCGCAACGAGGCCACUGAUGGAACCCUUAAGAUUCAAGUAUCUU UGCAGAUUGGCAUUAAGACAGAUGAUUCCCAUGACUGGACAAAGCUU CGGUACAUGGACUCACACACGCCUGCAGAUGCUGAAAGGGCAGGGCU CUUGGUCAGGACCUCGGCCCCUUGUACAAUUACCGGGACCAUGGGCC ACUUCAUCCUUGCACGCUGCCCUAAGGGGGAGACGCUGACGGUGGGC UUUACUGACUCGCGUAAGAUCUCACACACAUGUACACACCCUUUCCA CCACGAACCUCCAGUCAUAGGGAGAGAGAGAUUUCACUCUCGCCCAC AGCAUGGCAAAGAGCUGCCAUGCUCCACAUAUGUCCAGAGCACUGCU GCUACCGCUGAAGAAAUUGAGGUUCACAUGCCACCCGAUACACCAGA CCGUACUCUGAUGACCCAACAGAGCGGCAACGUGAAGAUUACCGUAA AUGGACAGACCGUGAGAUAUAAGUGCAACUGUGGUGGCUCCAAUGAG GGCUUAACAACAACGGAUAAGGUGAUUAACAAUUGCAAAAUAGAUCA GUGCCAUGCCGCAGUGACCAAUCACAAGAAUUGGCAAUACAACUCAC CCCUAGUGCCGAGGAACGCAGAACUAGGCGACAGGAAAGGGAAAAUC CAUAUACCCUUCCCCCUAGCAAAUGUGACCUGCCGAGUGCCCAAGGC CAGAAACCCCACGGUUACUUACGGCAAGAACCAGGUGACGAUGCUUU UGUACCCAGACCAUCCCACCUUGCUCUCUUAUAGAAACAUGGGACAG GAGCCUAACUAUCAUGAGGAGUGGGUGACACACAAGAAAGAAGUCAC CCUUACCGUGCCUACCGAAGGGCUUGAAGUCACCUGGGGCAACAACG AGCCUUACAAGUAUUGGCCACAGAUGUCCACAAACGGAACAGCCCAC GGCCACCCGCACGAGAUCAUACUGUAUUACUAUGAGCUUUAUCCCAC AAUGACUGUCGUAAUUGUGAGCGUUGCCAGCUUCGUGUUGCUUUCAA UGGUUGGCACUGCCGUGGGGAUGUGCGUGUGUGCUAGGCGCCGCUGU AUAACUCCUUAUGAACUAACUCCAGGCGCCACCGUUCCUUUCCUGCU CUCACUACUGUGUUGUGUGCGCACAACAAAGGCUGCCACCUACUACG AAGCCGCCGCCUACUUAUGGAAUGAACAGCAGCCUCUCUUUUGGUUA CAGGCGCUGAUUCCUCUUGCUGCCCUGAUCGUGCUAUGCAACUGCCU CAAGCUGCUGCCCUGUUGUUGCAAGACCCUAGCUUUUCUCGCCGUGA UGAGCAUCGGGGCACAUACAGUGUCCGCCUAUGAGCACGUCACCGUU AUCCCGAACACCGUCGGUGUGCCAUAUAAGACGUUAGUCAAUCGACC CGGCUACUCUCCAAUGGUGCUGGAAAUGGAACUCCAGAGUGUGACAC UGGAGCCAACCUUAUCCCUCGAUUAUAUUACCUGCGAAUACAAGACC GUCAUCCCUUCACCCUAUGUCAAGUGCUGUGGGACCGCUGAAUGCAA AGACAAGAGCUUGCCUGAUUACAGUUGCAAGGUCUUCACAGGUGUGU ACCCCUUCAUGUGGGGGGGAGCUUAUUGCUUUUGUGAUGCUGAGAAC ACCCAACUGAGCGAGGCUCACGUCGAGAAAUCUGAGUCUUGCAAGAC CGAGUUUGCCUCAGCUUACAGGGCCCACACGGCCAGCGCAUCCGCCA AAUUGAGGGUACUCUACCAGGGUAAUAAUAUCACCGUUGCCGCAUAU GCAAACGGCGAUCACGCCGUGACUGUCAAGGAUGCCAAGUUCGUUGU GGGCCCCAUGUCUAGCGCUUGGACACCGUUCGAUAAUAAGAUCGUCG UGUACAAAGGGGACGUGUAUAAUAUGGACUACCCACCUUUCGGGGCC GGCCGACCGGGACAGUUCGGGGAUAUUCAGAGCCGCACACCCGAAUC UAAAGAUGUUUACGCCAAUACUCAGCUCGUCCUGCAGAGGCCCGCCG CUGGUACAGUUCACGUUCCUUACUCACAGGCACCCUCUGGGUUUAAG UAUUGGCUGAAAGAACGAGGUGCCAGCUUGCAGCAUACAGCGCCUUU CGGAUGCCAGAUUGCCACUAACCCCGUACGGGCUGUCAACUGCGCGG UCGGCAAUAUUCCCAUUAGCAUUGAUAUCCCGGACGCAGCUUUCACC AGGGUUGUGGACGCCCCGAGCGUCACCGACAUGAGUUGUGAGGUGCC AGCCUGCACGCAUAGCAGUGAUUUCGGCGGCGUCGCCAUCAUUAAAU AUACCGCAAGCAAGAAAGGCAAGUGCGCCGUCCACUCGAUGACUAAC GCCGUCACAAUUCGGGAAGCCGAUGUUGAGGUCGAAGGCAACUCCCA GCUGCAGAUCAGCUUCUCUACUGCUCUUGCAAGCGCCGAGUUUCGAG UCCAGGUCUGCAGUACGCAGGUGCAUUGUGCAGCUGCCUGCCAUCCA CCCAAAGAUCAUAUUGUGAAUUAUCCGGCGUCACAUACCACACUGGG GGUCCAGGAUAUUAGUACAACGGCGAUGUCCUGGGUGCAGAAAAUUA CGGGAGGAGUGGGCUUAAUUGUUGCCGUGGCGGCCCUGAUCCUGAUC GUUGUGCUGUGUGUUAGCUUCUCUAGGCAUUGAUAAUAGGCUGGAGC CUCGGUGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCU CCCCUUCCUGCACCCGUACCCCCGUGGUCUUUGAAUAAAGUCUGAGU GGGCGGC

TABLE 48  CHIKV Polypeptide Sequences Antigen SEQ ID identifier Amino acid sequence NO: SE_chikv- MYEHVTVIPNTVGVPYKTLVNRPGYSPMVLEMELLSVTLEPTLSLDYITCEYK 402 Brazillian- TVIPSPYVKCCGTAECKDKSLPDYSCKVFTGVYPFMWGGAYCFCDTENTQLS E1_KP164567- EAHVEKSESCKTEFASAYRAHTASASAKLRVLYQGNNITVAAYANGDHAVT 71_72 VKDAKFIVGPMSSAWTPFDNKIVVYKGDVYNMDYPPFGAGRPGQFGDIQSRT PESEDVYANTQLVLQRPSAGTVHVPYSQAPSGFKYWLKERGASLQHTAPFGC QIATNPVRAMNCAVGNMPISIDIPDAAFTRVVDAPSLTDMSCEVSACTHSSDF GGVAIIKYAASKKGKCAVHSMTNAVTIREAEIEVEGNSQLQISFSTALASAEFR VQVCSTQVHCAAECHPPKDHIVNYPASHTTLGVQDISATAMSWVQKITGGVG LVVAVAALILIVVLCVSFSRH SE_chikv- MYEHVTVIPNTVGVPYKTLVNRPGYSPMVLEMELLSVTLEPTLSLDYITCEYK 403 Brazillian- TVIPSPYVKCCGTAECKDKNLPDYSCKVFTGVYPFMWGGAYCFCDAENTQLS E1_KP164568- EAHVEKSESCKTEFASAYRAHTASASAKLRVLYQGNNITVTAYANGDHAVT 69_70 VKDAKFIVGPMSSAWTPFDNKIVVYKGDVYNMDYPPFGAGRPGQFGDIQSRT PESKDVYANTQLVLQRPAAGTVHVPYSQAPSGFKYWLKERGASLQHTAPFG CQIATNPVRAVNCAVGNMPISIDIPDAAFIRVVDAPSLTDMSCEVPACTHSSDF GGVAIIKYAASKKGKCAVHSMTNAVTIREAEIEVEGNSQLQISFSTALASAEFR VQVCSTQVHCVAECHPPKDHIVNYPASHTTLGVQDISATALSWVQKITGGVG LVVAVAALILIVVLCVSFSRH SE_chikv- MSIKDHFNVYKATRPYLAHCPDCGEGHSCHSPVALERIRNEATDGTLKIQVSL 404 Brazillian- QIGIKTDDSHDWTKLRYMDNHMPADAERAGLFVRTSAPCTITGTMGHFILAR E2-E1_ CPKGETLTVGFTDGRKISHSCTHPFHHDPPVIGREKFHSRPQHGRELPCSTYAQ KP164567- STAATAEEIEVHMPPDTPDRTLMSQQSGNVKITVNSQTVRYKCNCGDSSEGLT 71_72 TTDKVINNCKVDQCHAAVTNHKKWQYNSPLVPRNAEFGDRKGKVHIPFPLA NVTCRVPKARNPTVTYGKNQVIMLLYPDHPTLLSYRNMGEEPNYQEEWVTH KKEIRLTVPTEGLEVTWGNNEPYKYWPQLSTNGTAHGHPHEIILYYYELYPT MTAVVLSVASFILLSMVGVAVGMCMCARRRCITPYELTPGATVPFLLSLICCI RTAKAYEHVTVIPNTVGVPYKTLVNRPGYSPMVLEMELLSVTLEPTLSLDYIT CEYKTVIPSPYVKCCGTAECKDKSLPDYSCKVFTGVYPFMWGGAYCFCDTEN TQLSEAHVEKSESCKTEFASAYRAHTASASAKLRVLYQGNNITVAAYANGDH AVTVKDAKFIVGPMSSAWTPFDNKIVVYKGDVYNMDYPPFGAGRPGQFGDI QSRTPESEDVYANTQLVLQRPSAGTVHVPYSQAPSGFKYWLKERGASLQHTA PFGCQIATNPVRAMNCAVGNMPISIDIPDAAFTRVVDAPSLTDMSCEVSACTH SSDFGGVAIIKYAASKKGKCAVHSMTNAVTIREAEIEVEGNSQLQISFSTALAS AEFRVQVCSTQVHCAAECHPPKDHIVNYPASHTTLGVQDISATAMSWVQKIT GGVGLVVAVAALILIVVLCVSFSRHMSIKDHFNVYKATRPYLAHCPDCGEGH SCHSPVALERIRNEATDGTLKIQVSLQIGIKTDDSHDWTKLRYMDNHMPADA ERAGLFVRTSAPCTITGTMGHFILARCPKGETLTVGFTDGRKISHSCTHPFHHD PPVIGREKFHSRPQHGRELPCSTYAQSTAATAEEIEVHMPPDTPDRTLMSQQS GNVKITVNSQTVRYKCNCGDSSEGLTTTDKVINNCKVDQCHAAVTNHKKWQ YNSPLVPRNAEFGDRKGKVHIPFPLANVTCRVPKARNPTVTYGKNQVIMLLY PDHPTLLSYRNMGEEPNYQEEWVTHKKEIRLTVPTEGLEVTWGNNEPYKYW PQLSTNGTAHGHPHEIILYYYELYPTMTAVVLSVASFILLSMVGVAVGMCMC ARRRCITPYELTPGATVPFLLSLICCIRTAKAYEHVTVIPNTVGVPYKTLVNRP GYSPMVLEMELLSVTLEPTLSLDYITCEYKTVIPSPYVKCCGTAECKDKSLPD YSCKVFTGVYPFMWGGAYCFCDTENTQLSEAHVEKSESCKTEFASAYRAHT ASASAKLRVLYQGNNITVAAYANGDHAVTVKDAKFIVGPMSSAWTPFDNKI VVYKGDVYNMDYPPFGAGRPGQFGDIQSRTPESEDVYANTQLVLQRPSAGTV HVPYSQAPSGFKYWLKERGASLQHTAPFGCQIATNPVRAMNCAVGNMPISIDI PDAAFTRVVDAPSLTDMSCEVSACTHSSDFGGVAIIKYAASKKGKCAVHSMT NAVTIREAEIEVEGNSQLQISFSTALASAEFRVQVCSTQVHCAAECHPPKDHIV NYPASHTTLGVQDISATAMSWVQKITGGVGLVVAVAALILIVVLCVSFSRH SQ-031495 MSTKDNFNVYKATRPYLAHCPDCGEGHSCHSPVALERIRNEATDGTLKIQVS 405 SE_chikv- LQIGIKTDDSHDWTKLRYMDNHTPADAERAGLFVRTSAPCTITGTMGHFILTR Brazillian- CPKGETLTVGFTDSRKISHSCTHPFHHDPPVIGREKFHSRPQHGKELPCSTYVQ E2-E1_ STAATTEEIEVHMPPDTPDRTLMSQQSGNVKITVNGQTVRYKCNCGGSNEGLI KP164568- TTDKVINNCKVDQCHAAVTNHKKWQYNSPLVPRNAELGDRKGKIHIPFPLAN 69_70 VTCRVPKARNPTVTYGKNQVIMLLYPDHPTLLSYRNMGEEPNYQEEWVTHK KEVVLTVPTEGLEVTWGNNEPYKYWPQLSTNGTAHGHPHEIILYYYELYPTM TVVVVSVASFVLLSMVGVAVGMCMCARRRCITPYELTPGATVPFLLSLICCIR TAKAYEHVTVIPNTVGVPYKTLVNRPGYSPMVLEMELLSVTLEPTLSLDYITC EYKTVIPSPYVKCCGTAECKDKNLPDYSCKVFTGVYPFMWGGAYCFCDAEN TQLSEAHVEKSESCKTEFASAYRAHTASASAKLRVLYQGNNITVTAYANGDH AVTVKDAKFIVGPMSSAWTPFDNKIVVYKGDVYNMDYPPFGAGRPGQFGDI QSRTPESKDVYANTQLVLQRPAAGTVHVPYSQAPSGFKYWLKERGASLQHT APFGCQIATNPVRAVNCAVGNMPISIDIPDAAFIRVVDAPSLTDMSCEVPACTH SSDFGGVAIIKYAASKKGKCAVHSMTNAVTIREAEIEVEGNSQLQISFSTALAS AEFRVQVCSTQVHCVAECHPPKDHIVNYPASHTTLGVQDISATALSWVQKIT GGVGLVVAVAALILIVVLCVSFSRH SE_chikv- MSIKDHFNVYKATRPYLAHCPDCGEGHSCHSPVALERIRNEATDGTLKIQVSL 406 Brazillian- QIGIKTDDSHDWTKLRYMDNHMPADAERAGLFVRTSAPCTITGTMGHFILAR E2_KP164567- CPKGETLTVGFTDGRKISHSCTHPFHHDPPVIGREKFHSRPQHGRELPCSTYAQ 71_72 STAATAEEIEVHMPPDTPDRTLMSQQSGNVKITVNSQTVRYKCNCGDSSEGLT TTDKVINNCKVDQCHAAVTNHKKWQYNSPLVPRNAEFGDRKGKVHIPFPLA NVTCRVPKARNPTVTYGKNQVIMLLYPDHPTLLSYRNMGEEPNYQEEWVTH KKEIRLTVPTEGLEVTWGNNEPYKYWPQLSTNGTAHGHPHEIILYYYELYPT MTAVVLSVASFILLSMVGVAVGMCMCARRRCITPYELTPGATVPFLLSLICCI RTAKA SE_chikv- MSTKDNFNVYKATRPYLAHCPDCGEGHSCHSPVALERIRNEATDGTLKIQVS 407 Brazillian- LQIGIKTDDSHDWTKLRYMDNHTPADAERAGLFVRTSAPCTITGTMGHFILTR E2_KP164568- CPKGETLTVGFTDSRKISHSCTHPFHHDPPVIGREKFHSRPQHGKELPCSTYVQ 69_70 STAATTEEIEVHMPPDTPDRTLMSQQSGNVKITVNGQTVRYKCNCGGSNEGLI TTDKVINNCKVDQCHAAVTNHKKWQYNSPLVPRNAELGDRKGKIHIPFPLAN VTCRVPKARNPTVTYGKNQVIMLLYPDHPTLLSYRNMGEEPNYQEEWVTHK KEVVLTVPTEGLEVTWGNNEPYKYWPQLSTNGTAHGHPHEIILYYYELYPTM TVVVVSVASFVLLSMVGVAVGMCMCARRRCITPYELTPGATVPFLLSLICCIR TAKA SE_CHIKV_ MEFIPTQTFYNRRYQPRPWAPRPTIQVIRPRPRPQRQAGQLAQLISAVNKLTM 408 C_E3_E2_6K_ RAVPQQKPRRNRKNKKQRQKKQAPQNDPKQKKQPPQKKPAQKKKKPGRRE E1_no Flag RMCMKIENDCIFEVKHEGKVMGYACLVGDKVMKPAHVKGTIDNADLAKLA or V5 or HA FKRSSKYDLECAQIPVHMKSDASKFTHEKPEGYYNWHHGAVQYSGGRFTIPT (Strain 37997 GAGKPGDSGRPIFDNKGRVVAIVLGGANEGARTALSVVTWNKDIVTKITPEG Senegal) AEEWSLALPVLCLLANTTFPCSQPPCTPCCYEKEPESTLRMLEDNVMRPGYYQ LLKASLTCSPHRQRRSTKDNFNVYKATRPYLAHCPDCGEGHSCHSPIALERIR NEATDGTLKIQVSLQIGIKTDDSHDWTKLRYMDSHTPADAERAGLLVRTSAP CTITGTMGHFILARCPKGETLTVGFTDSRKISHTCTHPFHHEPPVIGRERFHSRP QHGKELPCSTYVQSTAATAEEIEVHMPPDTPDRTLMTQQSGNVKITVNGQTV RYKCNCGGSNEGLTTTDKVINNCKIDQCHAAVTNHKNWQYNSPLVPRNAEL GDRKGKIHIPFPLANVTCRVPKARNPTVTYGKNQVTMLLYPDHPTLLSYRNM GQEPNYHEEWVTHKKEVTLTVPTEGLEVTWGNNEPYKYWPQMSTNGTAHG HPHEIILYYYELYPTMTVVIVSVASFVLLSMVGTAVGMCVCARRRCITPYELT PGATVPFLLSLLCCVRTTKAATYYEAAAYLWNEQQPLFWLQALIPLAALIVLC NCLKLLPCCCKTLAFLAVMSIGAHTVSAYEHVTVIPNTVGVPYKTLVNRPGYS PMVLEMELQSVTLEPTLSLDYITCEYKTVIPSPYVKCCGTAECKDKSLPDYSC KVFTGVYPFMWGGAYCFCDAENTQLSEAHVEKSESCKTEFASAYRAHTASA SAKLRVLYQGNNITVAAYANGDHAVTVKDAKFVVGPMSSAWTPFDNKIVVY KGDVYNMDYPPFGAGRPGQFGDIQSRTPESKDVYANTQLVLQRPAAGTVHV PYSQAPSGFKYWLKERGASLQHTAPFGCQIATNPVRAVNCAVGNIPISIDIPDA AFTRVVDAPSVTDMSCEVPACTHSSDFGGVAIIKYTASKKGKCAVHSMTNAV TIREADVEVEGNSQLQISFSTALASAEFRVQVCSTQVHCAAACHPPKDHIVNY PASHTTLGVQDISTTAMSWVQKITGGVGLIVAVAALILIVVLCVSFSRH SE_CHIKV_ MEFIPTQTFYNRRYQPRPWAPRPTIQVIRPRPRPQRQAGQLAQLISAVNKLTM 409 C_E3_E2_6K_ RAVPQQKPRRNRKNKKQRQKKQAPQNDPKQKKQPPQKKPAQKKKKPGRRE E1_no Flag RMCMKIENDCIFEVKHEGKVMGYACLVGDKVMKPAHVKGTIDNADLAKLA or V5 or FKRSSKYDLECAQIPVHMKSDASKFTHEKPEGYYNWHHGAVQYSGGRFTIPT HA_DX GAGKPGDSGRPIFDNKGRVVAIVLGGANEGARTALSVVTWNKDIVTKITPEG AEEWSLALPVLCLLANTTFPCSQPPCTPCCYEKEPESTLRMLEDNVMRPGYYQ LLKASLTCSPHRQRRSTKDNFNVYKATRPYLAHCPDCGEGHSCHSPIALERIR NEATDGTLKIQVSLQIGIKTDDSHDWTKLRYMDSHTPADAERAGLLVRTSAP CTITGTMGHFILARCPKGETLTVGFTDSRKISHTCTHPFHHEPPVIGRERFHSRP QHGKELPCSTYVQSTAATAEEIEVHMPPDTPDRTLMTQQSGNVKITVNGQTV RYKCNCGGSNEGLTTTDKVINNCKIDQCHAAVTNHKNWQYNSPLVPRNAEL GDRKGKIHIPFPLANVTCRVPKARNPTVTYGKNQVTMLLYPDHPTLLSYRNM GQEPNYHEEWVTHKKEVTLTVPTEGLEVTWGNNEPYKYWPQMSTNGTAHG HPHEIILYYYELYPTMTVVIVSVASFVLLSMVGTAVGMCVCARRRCITPYELT PGATVPFLLSLLCCVRTTKAATYYEAAAYLWNEQQPLFWLQALIPLAALIVLC NCLKLLPCCCKTLAFLAVMSIGAHTVSAYEHVTVIPNTVGVPYKTLVNRPGYS PMVLEMELQSVTLEPTLSLDYITCEYKTVIPSPYVKCCGTAECKDKSLPDYSC KVFTGVYPFMWGGAYCFCDAENTQLSEAHVEKSESCKTEFASAYRAHTASA SAKLRVLYQGNNITVAAYANGDHAVTVKDAKFVVGPMSSAWTPFDNKIVVY KGDVYNMDYPPFGAGRPGQFGDIQSRTPESKDVYANTQLVLQRPAAGTVHV PYSQAPSGFKYWLKERGASLQHTAPFGCQIATNPVRAVNCAVGNIPISIDIPDA AFTRVVDAPSVTDMSCEVPACTHSSDFGGVAIIKYTASKKGKCAVHSMTNAV TIREADVEVEGNSQLQISFSTALASAEFRVQVCSTQVHCAAACHPPKDHIVNY PASHTTLGVQDISTTAMSWVQKITGGVGLIVAVAALILIVVLCVSFSRH SE_CHIKV_E1_ MYEHVTVIPNTVGVPYKTLVNRPGYSPMVLEMELQSVTLEPTLSLDYITCEYK 410 no Flag or TVIPSPYVKCCGTAECKDKSLPDYSCKVFTGVYPFMWGGAYCFCDAENTQLS V5 EAHVEKSESCKTEFASAYRAHTASASAKLRVLYQGNNITVAAYANGDHAVT VKDAKFVVGPMSSAWTPFDNKIVVYKGDVYNMDYPPFGAGRPGQFGDIQSR TPESKDVYANTQLVLQRPAAGTVHVPYSQAPSGFKYWLKERGASLQHTAPFG CQIATNPVRAVNCAVGNIPISIDIPDAAFTRVVDAPSVTDMSCEVPACTHSSDF GGVAIIKYTASKKGKCAVHSMTNAVTIREADVEVEGNSQLQISFSTALASAEF RVQVCSTQVHCAAACHPPKDHIVNYPASHTTLGVQDISTTAMSWVQKITGGV GLIVAVAALILIVVLCVSFSRH CHIKV_E2_6K_ MSTKDNFNVYKATRPYLAHCPDCGEGHSCHSPIALERIRNEATDGTLKIQVSL 411 E1_no Flag QIGIKTDDSHDWTKLRYMDSHTPADAERAGLLVRTSAPCTITGTMGHFILARC or V5 PKGETLTVGFTDSRKISHTCTHPFHHEPPVIGRERFHSRPQHGKELPCSTYVQS TAATAEEIEVHMPPDTPDRTLMTQQSGNVKITVNGQTVRYKCNCGGSNEGLT TTDKVINNCKIDQCHAAVTNHKNWQYNSPLVPRNAELGDRKGKIHIPFPLAN VTCRVPKARNPTVTYGKNQVTMLLYPDHPTLLSYRNMGQEPNYHEEWVTHK KEVTLTVPTEGLEVTWGNNEPYKYWPQMSTNGTAHGHPHEIILYYYELYPTM TVVIVSVASFVLLSMVGTAVGMCVCARRRCITPYELTPGATVPFLLSLLCCVR TTKAATYYEAAAYLWNEQQPLFWLQALIPLAALIVLCNCLKLLPCCCKTLAF LAVMSIGAHTVSAYEHVTVIPNTVGVPYKTLVNRPGYSPMVLEMELQSVTLE PTLSLDYITCEYKTVIPSPYVKCCGTAECKDKSLPDYSCKVFTGVYPFMWGGA YCFCDAENTQLSEAHVEKSESCKTEFASAYRAHTASASAKLRVLYQGNNITV AAYANGDHAVTVKDAKFVVGPMSSAWTPFDNKIVVYKGDVYNMDYPPFGA GRPGQFGDIQSRTPESKDVYANTQLVLQRPAAGTVHVPYSQAPSGFKYWLKE RGASLQHTAPFGCQIATNPVRAVNCAVGNIPISIDIPDAAFTRVVDAPSVTDMS CEVPACTHSSDFGGVAIIKYTASKKGKCAVHSMTNAVTIREADVEVEGNSQL QISFSTALASAEFRVQVCSTQVHCAAACHPPKDHIVNYPASHTTLGVQDISTT AMSWVQKITGGVGLIVAVAALILIVVLCVSFSRH SE_CHIKV_ MSTKDNFNVYKATRPYLAHCPDCGEGHSCHSPIALERIRNEATDGTLKIQVSL 412 E2_no Flag QIGIKTDDSHDWTKLRYMDSHTPADAERAGLLVRTSAPCTITGTMGHFILARC or V5 PKGETLTVGFTDSRKISHTCTHPFHHEPPVIGRERFHSRPQHGKELPCSTYVQS TAATAEEIEVHMPPDTPDRTLMTQQSGNVKITVNGQTVRYKCNCGGSNEGLT TTDKVINNCKIDQCHAAVTNHKNWQYNSPLVPRNAELGDRKGKIHIPFPLAN VTCRVPKARNPTVTYGKNQVTMLLYPDHPTLLSYRNMGQEPNYHEEWVTHK KEVTLTVPTEGLEVTWGNNEPYKYWPQMSTNGTAHGHPHEIILYYYELYPTM TVVIVSVASFVLLSMVGTAVGMCVCARRRCITPYELTPGATVPFLLSLLCCVR TTKA CHIKVC-E3- SSFWTLVQKLIRLTIGKERKEEEEIEPPWSLSLRRRSIIGGTSPGLGPPALQSKDH 413 E2-6K-E1 VPGRRDKPANWRNSSAQLTSPCERFLSRSRGGTGRIRNNAKRNRRRRTTLNR RNNLPRKSQLRRRRSLDAVKECAKSKMIASLRSTRAKWGTHAWWAIRSSQH MRGQSIMLIWPSLLNVAPNTILSVPRYLCTNLMQASSHTRSLRAIITGIMVRFS TPAADLPFLQGQESRAIRGDPFSTTKAAWLSCSVGLMRVHVLHLAWLPGIRTL SQRLHRREQRNGAWHCPFCACWPTPLSHVVNPLALPAAMRKSLRARYVCW KIMSGPGTINCSRLVHARPTGSAGPRKITSTFTRLLGLIWPTVPIAERDILVIVLL PWSESATRPLMEPLRFKYLCRLALRQMIPMTGQSFGTWTHTRLQMLKGQGS WSGPRPLVQLPGPWATSSLHAALRGRRRWALLTRVRSHTHVHTLSTTNLQSG ERDFTLAHSMAKSCHAPHMSRALLLPLKKLRFTCHPIHQTVLPNRAATRLPM DRPDISATVVAPMRAQQRIRLTIAKISAMPQPITRIGNTTHPCRGTQNATGKGK SIYPSPQMPAECPRPETPRLLTARTRRCFCTQTIPPCSLIETWDRSLTIMRSGHT RIU(SPLPCLPKGLKSPGATTSLTSIGHRCPQTEQPTATRTRSYCITMSFIPQLSL ALPASCCFQWLALPWGCACVLGAAVLLMNLQAPPFLSCSHYCVVCAQQRLP PTTKPPPTYGMNSSLSFGYRRFLLLPSCYATASSCCPVVARPLFSPASGHIQCPP MSTSPLSRTPSVCHIRRSIDPATLQWCWKWNSRVHWSQPYPSIILPANTRPSSL HPMSSAVGPLNAKTRACLITVARSSQVCTPSCGGELIAFVMLRTPNARLTSRN LSLARPSLPQLTGPTRPAHPPNGYSTRVIISPLPHMQTAITPLSRMPSSLWAPCL ALGHRSIIRSSCTKGTCIIWTTHLSGPADRDSSGIFRAAHPNLKMFTPILSSSCRG PPLVQFTFLTHRHPLGLSIGKNEVPACSIQRLSDARLPLTPYGLSTARSAIFPLA LISRTQLSPGLWTPRASPTVVRCQPARIAVISAASPSLNIPQARKASAPSTRLTP SQFGKPMLRSKATPSCRSASLLLLQAPSFESRSAVRRCIVQLPAIHPKIILIIRRH IPHWGSRILVQRRCPGCRKLREEWALLPWRPSSLCCVLASLGIDNRLEPRWPCF LPLGPPPSPSSPSCTRTPVVFESLSGR

TABLE 49 Study design, schedule of injection/bleeding, readout, and survival data for the 2 μg dose study of the CHIKV E1, CHIKV E2, and CHIKV E1/E2/E3/6K/C vaccines Day 6 post- Group Vaccine injection (n = 5) Vaccine Schedule Dosage/Route Challenge Readout (survival/total) 1 E1 Day 0 IM, LNP Challenge Retro-orbital 0/5 2 Day 0, 28 2 μg with 1g4 PFU bleeds on 0/5 3 Day 0 ID, LNP per mouse of Days 7, 28, 0/5 4 Day 0, 28 2 μg CHIK 181/25 56. 0/5 5 E2 Day 0 IM, LNP via ID Serum of 0/5 6 Day 0, 28 2 μg injection mice that 4/5 7 Day 0 ID, LNP (Day 56). survive 0/5 8 Day 0, 28 2 μg Weights and challenge 5/5 9 E1/E2/E3 C Day 0 IM, LNP health for 10 will undergo 5/5 10 Day 0, 28 2 μg days E1 & E2 5/5 11 Day 0 ID, LNP ELISA 4/5 12 Day 0, 28 2 μg testing. 5/5 13 HI CHIRV (+) Day 0 IN, 20 ul 0/5 14 Day 0, 28 0/5 15 (−) 0/5

TABLE 50 Study design, schedule of injection/bleeding, readout, and survival data for the 10 μg dose study of the CHIKV E1, CHIKV E2, and CHIKV C-E3-E2-6K-E1 vaccines Day 6 post- Group Vaccine intection (n = 5) Vaccine Schedule Dosage/Route Challenge Readout (survivor/total) 1 E1 Day 0 IM, LNP Challenge Retro-orbital 0/5 2 Day 0, 28 10 μg with 1e4 PFU bleeds on 3/5 3 Day 0 ID, LNP per mouse of Days 7, 28, 0/5 4 Day 0, 28 10 μg CHIK 181/25 56. 4/5 5 E2 Day 0 IM, LNP via ID Serum of 0/5 6 Day 0, 28 10 μg injection mice that 5/5 7 Day 0 ID, LNP (Day 56). survive 0/5 8 Day 0, 28 10 μg Weights and challenge 5/5 9 E1/E2/E3 C Day 0 IM, LNP health for 10 will undergo 5/5 10 Day 0, 28 10 μg days E1 & E2 5/5 11 Day 0 ID, LNP ELISA 5/5 12 Day 0, 28 10 μg testing. 5/5 13 HI CHIKV (+) Day 0 IN, 20 ul 0/5 14 Day 0, 28 0/5 15 (−) 0/5

TABLE 51 Health Status SCORE INITIALS DESCRIPTION APPEARANCE MOBILITY ATTITUDE 1 H Healthy Smooth Coat. Active, Alert Bright Eyes. Scurrying,

2 SR Slightly Ruffled Slightly Ruffled Active, Alert coat (usually only Scurrying, around head and Burrowing 3 R Ruffled Ruffled Coat Active, Alert throughout body. Scurrying, A “wet” Burrowing 4 S Sick Very Ruffled coat. Walking, but no Mildly Slightly closed, scurrying. Lethargic inset eyes. 5 VS Very Sick Very Ruffled Slow to no Extremely (Euthanize) Coat. Closed, inset movement. Will Lethargic 6 E Euthanize Very ruffled Coat. No movement or Completely Closed, inset eyes. Uncontrollable, Unaware or in Moribund spastic Noticeable requiring humane movements. Will Distress euthanasia. NOT return to upright position 7 D Deceased — — —

indicates data missing or illegible when filed

TABLE 52 Survival of mice vaccinated with Chikungunya E1 antigen mRNA - 2 μg dose E1 IM days post LNP E1 IM LNP E1 ID LNP E1 ID LNP infection Day 0 Day 0, 28 Day 0 Day 0, 28 Vehicle 0.000 100.000 100.000 100.000 100.000 100.000 4.000 80.000 40.000 40.000 60.000 5.000 0.000 0.000 0.000 0.000 0.000

TABLE 53 Survival of mice vaccinated with Chikungunya E1 antigen mRNA - 10 μg dose E1 IM days post LNP E1 IM LNP E1 ID LNP E1 ID LNP infection Day 0 Day 0, 28 Day 0 Day 0, 28 Vehicle 0.000 100.000 100.000 100.000 100.000 100.000 4.000 60.000 80.000 5.000 0.000 80.000 0.000 0.000 6.000 60.000 80.000 10.000 60.000 80.000

TABLE 54 Survival of mice vaccinated with Chikungunya E2 antigen mRNA - 2 μg dose E1 IM days post LNP E1 IM LNP E1 ID LNP E1 ID LNP infection Day 0 Day 0, 28 Day 0 Day 0, 28 Vehicle 0.000 100.000 100.000 100.000 100.000 100.000 3.000 60.000 4.000 20.000 80.000 0.000 5.000 0.000 0.000 0.000 6.000 80.000 10.000 80.000 100.000

TABLE 55 Survival of mice vaccinated with Chikungunya E2 antigen mRNA - 10 μg dose E2 IM days post LNP E2 IM LNP E2 ID LNP E2 ID LNP infection Day 0 Day 0, 28 Day 0 Day 0, 28 Vehicle 0.000 100.000 100.000 100.000 100.000 100.000 5.000 40.000 0.000 0.000 6.000 0.000 10.000 100.000 100.000

TABLE 56 Survival of mice vaccinated with Chikungunya C-E3-E2-6K-E1 antigen mRNA - 2 μg E1/E2/ E3C E1/E2/E3C E1/E2/E3C E1/E2/E3C days post IM LNP IM LNP ID LNP ID LNP infection Day 0 Day 0, 28 Day 0 Day 0, 28 Vehicle 0.000 100.000 100.000 100.000 100.000 100.000 5.000 80.000 0.000 10.000 100.000 100.000 80.000 100.000

TABLE 57 Survival of mice vaccinated with Chikungunya C-E3-E2-6K-E1 antigen mRNA - 10 μg E1/E2/ E3C E1/E2/E3C E1/E2/E3C E1/E2/E3C days post IM LNP IM LNP ID LNP ID LNP infection Day 0 Day 0, 28 Day 0 Day 0, 28 Vehicle 0.000 100.000 100.000 100.000 100.000 100.000 5.000 0.000 10.000 100.000 100.000 100.000 100.000

TABLE 58 Summary of Day 6 post-injection survival data Dose Dose 10 ug/mouse 2 ug/mouse G# Antigen/route/regime (survival %) (survival %) 1 Chik-E1-IM-single dose 0 0 2 Chik-E1-IM-two doses 60 0 3 Chik-E1-ID-single dose 0 0 4 Chik-E1-ID-two doses 80 0 5 Chik-E2-IM-single dose 0 0 6 Chik-E2-IM-two doses 100 80 7 Chik-E2-ID-single dose 0 0 8 Chik-E2-ID-two doses 100 100 9 Chik-E1-E2-E3-C-6KIM-single dose 100 100 10 Chik-E1-E2-E3-C-6KIM-two doses 100 100 11 Chik-E1-E2-E3-C-6KID-single dose 100 80 12 Chik-E1-E2-E3-C-6KID-two doses 100 100 13 HI CHIKV (+) 0 0 14 HI CHIKV (+) 0 0 15 Control (−) 0 0

TABLE 59 CHIKV Challenge Study Design in AG129 mice Group* Dose (n = 5) Vaccine Schedule (IM) Challenge Bleeds 1 VAL-181388 Day 0 10 μg Challenge with Pre-bleed for serum 2 Day 0 & 28 1 × 10⁴ PFU per via RO route on 3 Day 0  2 μg mouse of CHIK days −3, 28, 56, (all 4 Day 0 & 28 181/25 via ID groups) & 84, 112 7 Day 0 0.4 μg  injection (groups 10-16 only). 8 Day 0 & 28 on day 56. Terminal bleed 9 PBS Day 0 & 28 — Weights and health surviving mice on for 10 days day 10 post following challenge. infection. Serum stored at 10 VAL-181388 Day 0 10 μg Challenge with −80° C. 11 Day 0 & 28 1 × 10⁴ PFU per 12 Day 0  2 μg mouse of CHIK 13 Day 0 & 28 181/25 via ID 14 Day 0 0.4 μg  injection on day 15 Day 0 & 28 112. Weights and 16 PBS Day 0 & 28 — health for 10 days following infection. *No group 5 or 6 in this study

TABLE 60 Equipment and Software Item Vendor Cat#/Model Syringes BD Various Animal Housing InnoVive Various Scale Ohause AV2101 Prism software GraphPad N/A Microplate Washer BioTek ELx405 Plate reader with SoftMax Pro Molecular Devices VersaMax version 5.4.5

TABLE 61 ELISA Reagents Storage Name Supplier cat# Temperature Notes DPBS 1X, sterile Corning 21-031- Ambient For dilution of coating antigen CM or equivalent StartingBlock T20 Thermo Scientific 2-8° C. For blocking non-specific (PBS) Blocking 37539 binding and use as diluent of Buffer Standards, unknown test sera and detection antibody SureBlue Reserve KPL 53-00-02 or 2-8° C. N/A TMB Microwell equivalent Peroxidase Substrate (1- Component) DPBS powder, non- Corning 55-031-PB 2-8° C. Use deionized water to sterile or equivalent dissolved DPBS powder from one bottle to a final volume of 10 liters of 1X DPBS TWEEN-20 Sigma-Aldrich Ambient Add 5 mL TWEEN-20 to 10 P1379-500ML or liters of 1X DPBS and mix equivalent well to prepare DPBS + 0.05% TWEEN-20 Wash Buffer for automatic plate washer

TABLE 62 Test Conditions Critical Reagent Please note: Coating antigens and standards Supplier cat# Storage are stored as single-use aliquots. and/or lot# Temperature Assay Parameter Coating antigens CHIKV recombinant E1 glycoprotein, IBT Bioservices, −70° C. or below 400 ng/well expressed in 293 mammalian cells IBT's lot Aug. 11, 2015 BCA = 0.351 mg/mL CHIKV recombinant E2 glycoprotein, ImmunoDx, cat# −70° C. or below 400 ng/well expressed in E. coli IBT's BCA = 1.291 mg/mL 80002, lot 10MY4 CHIKV 181/25 lysate from sucrose- IBT Bioservices, −70° C. or below 300 ng/well purified viruses, lysed by sonication lot Nov. 23, 2015 IBT's BCA = 1.316 mg/mL Standards Anti-E1 positive control Pooled mouse IBT Bioservices −70° C. or below Assigned, 30,812 serum from survivors of BS-1842 group Antibody Units/mL 4 (vaccinated with E1 mRNA 10 μg, ID, against E1 protein LNP on study days 0 and 28) day 66 terminal bleeds (10 days after CHIKV infection) Anti-E2 positive control, Pooled mouse IBT Bioservices −70° C. or below Assigned, 16912 serum from survivors of BS-1842 group Antibody Units/mL 8 (vaccinated with E2 mRNA 10 μg, ID, against E2 protein LNP on study days 0 and 28) day 66 Assigned 14,200 terminal bleeds (10 days after CHIKV Antibody Units/mL infection) Detection antibody Anti-mouse IgG (H + L)-HRP KPL, cat# 474- 2-8° C. 1:6000 dilution 1806, lot 140081

TABLE 63 Survival Percentage Groups 1-4 and 7-9, Day 56 Challenge Days 10 μg 2 μg 2 μg 0.4 μg 0.4 μg p.i. 10 μg Day 0 Day 0 & 28 Day 0 Day 0 & 28 Day 0 Day 0 & 28 PBS 0 100 100 100 100 100 100 100 3 80 4 0 40 80 5 0 0 10  100 100 100 100 Groups 10-16, Day 112 Challenge Days 10 μg 2 μg 2 μg 0.04 μg 0.04 μg p.i. 10 μg Day 0 Day 0 & 28 Day 0 Day 0 & 28 Day 0 Day 0 & 28 PBS 0 100 100 100 100 100 100 100 3 80 80 4 20 20 50 5 0 0 0 10  100 100 100 100

TABLE 64 CHIKV Plaque Reduction Neutralization Test (PRNT) Serum dilutions from 1/20 to 1/40960 Expt info Vaccination CHIKV strain GP# regimen 37997 sample ID PRNT80 titer PRNT50 titer 10 Day 0, CHIKV 37997 1  1/160  1/640 IM/10 μg working stock 2  1/320  1/320 titer = 3  1/160  1/640 780 PFU/ml 4  1/160   1/1280 5  1/320   1/1280 11 Day0/Day28, 1  1/640   1/2560 IM/10 μg 2   1/1280   1/1280 3  1/320   1/2560 4  1/640   1/5120 5   1/1280   1/5120 12 Day 0, IM/2 μg 1  1/20  1/80 2  1/40  1/320 3 <1/20  1/160 PRNT80 cutoff 4 <1/20  1/160 8 PFU 5 <1/20  1/20 13 Day0, Day28, 1  1/80  1/320 IM/2 μg 2  1/80  1/640 3  1/20  1/320 4  1/20  1/320 5  1/320  1/640 14 Day 0, 1 <1/20 80 IM/0.4 μg 2 <1/20 <1/20 3 <1/20 <1/20 4 <1/20 <1/20 5 <1/20 <1/20 15 Day0, Day28, PRNT50 cutoff 1 <1/20 <1/20 IM/0.4 μg 20 PFU 2 <1/20 80 3 <1/20 <1/20 4 <1/20 <1/20 5 <1/20 <1/20 16 Vehicle 1 <1/20 <1/20 Day0/Day28 2 <1/20 <1/20 3 <1/20 <1/20 4 <1/20 <1/20 5 <1/20 <1/20

TABLE 65  Flagellin Nucleic Acid Sequences SEQ ID Name Sequence NO: NT (5′ TCAAGCTTTTGGACCCTCGTACAGAAGCTAATACGACTCACTATAGGGA 417 UTR, ORF, AATAAGAGAGAAAAGAAGAGTAAGAAGAAATATAAGAGCCACCATGG 3′ UTR) CACAAGTCATTAATACAAACAGCCTGTCGCTGTTGACCCAGAATAACCT GAACAAATCCCAGTCCGCACTGGGCACTGCTATCGAGCGTTTGTCTTCC GGTCTGCGTATCAACAGCGCGAAAGACGATGCGGCAGGACAGGCGATT GCTAACCGTTTTACCGCGAACATCAAAGGTCTGACTCAGGCTTCCCGTA ACGCTAACGACGGTATCTCCATTGCGCAGACCACTGAAGGCGCGCTGA ACGAAATCAACAACAACCTGCAGCGTGTGCGTGAACTGGCGGTTCAGT CTGCGAATGGTACTAACTCCCAGTCTGACCTCGACTCCATCCAGGCTGA AATCACCCAGCGCCTGAACGAAATCGACCGTGTATCCGGCCAGACTCA GTTCAACGGCGTGAAAGTCCTGGCGCAGGACAACACCCTGACCATCCA GGTTGGTGCCAACGACGGTGAAACTATCGATATTGATTTAAAAGAAAT CAGCTCTAAAACACTGGGACTTGATAAGCTTAATGTCCAAGATGCCTAC ACCCCGAAAGAAACTGCTGTAACCGTTGATAAAACTACCTATAAAAAT GGTACAGATCCTATTACAGCCCAGAGCAATACTGATATCCAAACTGCA ATTGGCGGTGGTGCAACGGGGGTTACTGGGGCTGATATCAAATTTAAA GATGGTCAATACTATTTAGATGTTAAAGGCGGTGCTTCTGCTGGTGTTT ATAAAGCCACTTATGATGAAACTACAAAGAAAGTTAATATTGATACGA CTGATAAAACTCCGTTGGCAACTGCGGAAGCTACAGCTATTCGGGGAA CGGCCACTATAACCCACAACCAAATTGCTGAAGTAACAAAAGAGGGTG TTGATACGACCACAGTTGCGGCTCAACTTGCTGCAGCAGGGGTTACTGG CGCCGATAAGGACAATACTAGCCTTGTAAAACTATCGTTTGAGGATAA AAACGGTAAGGTTATTGATGGTGGCTATGCAGTGAAAATGGGCGACGA TTTCTATGCCGCTACATATGATGAGAAAACAGGTGCAATTACTGCTAAA ACCACTACTTATACAGATGGTACTGGCGTTGCTCAAACTGGAGCTGTGA AATTTGGTGGCGCAAATGGTAAATCTGAAGTTGTTACTGCTACCGATGG TAAGACTTACTTAGCAAGCGACCTTGACAAACATAACTTCAGAACAGG CGGTGAGCTTAAAGAGGTTAATACAGATAAGACTGAAAACCCACTGCA GAAAATTGATGCTGCCTTGGCACAGGTTGATACACTTCGTTCTGACCTG GGTGCGGTTCAGAACCGTTTCAACTCCGCTATCACCAACCTGGGCAATA CCGTAAATAACCTGTCTTCTGCCCGTAGCCGTATCGAAGATTCCGACTA CGCAACCGAAGTCTCCAACATGTCTCGCGCGCAGATTCTGCAGCAGGC CGGTACCTCCGTTCTGGCGCAGGCGAACCAGGTTCCGCAAAACGTCCTC TCTTTACTGCGTTGATAATAGGCTGGAGCCTCGGTGGCCATGCTTCTTG CCCCTTGGGCCTCCCCCCAGCCCCTCCTCCCCTTCCTGCACCCGTACCCC CGTGGTCTTTGAATAAAGTCTGAGTGGGCGGC ORF ATGGCACAAGTCATTAATACAAACAGCCTGTCGCTGTTGACCCAGAAT 418 Sequence, AACCTGAACAAATCCCAGTCCGCACTGGGCACTGCTATCGAGCGTTTGT NT CTTCCGGTCTGCGTATCAACAGCGCGAAAGACGATGCGGCAGGACAGG CGATTGCTAACCGTTTTACCGCGAACATCAAAGGTCTGACTCAGGCTTC CCGTAACGCTAACGACGGTATCTCCATTGCGCAGACCACTGAAGGCGC GCTGAACGAAATCAACAACAACCTGCAGCGTGTGCGTGAACTGGCGGT TCAGTCTGCGAATGGTACTAACTCCCAGTCTGACCTCGACTCCATCCAG GCTGAAATCACCCAGCGCCTGAACGAAATCGACCGTGTATCCGGCCAG ACTCAGTTCAACGGCGTGAAAGTCCTGGCGCAGGACAACACCCTGACC ATCCAGGTTGGTGCCAACGACGGTGAAACTATCGATATTGATTTAAAA GAAATCAGCTCTAAAACACTGGGACTTGATAAGCTTAATGTCCAAGAT GCCTACACCCCGAAAGAAACTGCTGTAACCGTTGATAAAACTACCTAT AAAAATGGTACAGATCCTATTACAGCCCAGAGCAATACTGATATCCAA ACTGCAATTGGCGGTGGTGCAACGGGGGTTACTGGGGCTGATATCAAA TTTAAAGATGGTCAATACTATTTAGATGTTAAAGGCGGTGCTTCTGCTG GTGTTTATAAAGCCACTTATGATGAAACTACAAAGAAAGTTAATATTGA TACGACTGATAAAACTCCGTTGGCAACTGCGGAAGCTACAGCTATTCG GGGAACGGCCACTATAACCCACAACCAAATTGCTGAAGTAACAAAAGA GGGTGTTGATACGACCACAGTTGCGGCTCAACTTGCTGCAGCAGGGGTT ACTGGCGCCGATAAGGACAATACTAGCCTTGTAAAACTATCGTTTGAG GATAAAAACGGTAAGGTTATTGATGGTGGCTATGCAGTGAAAATGGGC GACGATTTCTATGCCGCTACATATGATGAGAAAACAGGTGCAATTACTG CTAAAACCACTACTTATACAGATGGTACTGGCGTTGCTCAAACTGGAGC TGTGAAATTTGGTGGCGCAAATGGTAAATCTGAAGTTGTTACTGCTACC GATGGTAAGACTTACTTAGCAAGCGACCTTGACAAACATAACTTCAGA ACAGGCGGTGAGCTTAAAGAGGTTAATACAGATAAGACTGAAAACCCA CTGCAGAAAATTGATGCTGCCTTGGCACAGGTTGATACACTTCGTTCTG ACCTGGGTGCGGTTCAGAACCGTTTCAACTCCGCTATCACCAACCTGGG CAATACCGTAAATAACCTGTCTTCTGCCCGTAGCCGTATCGAAGATTCC GACTACGCAACCGAAGTCTCCAACATGTCTCGCGCGCAGATTCTGCAGC AGGCCGGTACCTCCGTTCTGGCGCAGGCGAACCAGGTTCCGCAAAACG TCCTCTCTTTACTGCGT mRNA G*GGGAAAUAAGAGAGAAAAGAAGAGUAAGAAGAAAUAUAAGAGCC 419 Sequence ACCAUGGCACAAGUCAUUAAUACAAACAGCCUGUCGCUGUUGACCCA (assumes  GAAUAACCUGAACAAAUCCCAGUCCGCACUGGGCACUGCUAUCGAGC T100 tail) GUUUGUCUUCCGGUCUGCGUAUCAACAGCGCGAAAGACGAUGCGGCA GGACAGGCGAUUGCUAACCGUUUUACCGCGAACAUCAAAGGUCUGAC UCAGGCUUCCCGUAACGCUAACGACGGUAUCUCCAUUGCGCAGACCA CUGAAGGCGCGCUGAACGAAAUCAACAACAACCUGCAGCGUGUGCGU GAACUGGCGGUUCAGUCUGCGAAUGGUACUAACUCCCAGUCUGACCU CGACUCCAUCCAGGCUGAAAUCACCCAGCGCCUGAACGAAAUCGACC GUGUAUCCGGCCAGACUCAGUUCAACGGCGUGAAAGUCCUGGCGCAG GACAACACCCUGACCAUCCAGGUUGGUGCCAACGACGGUGAAACUAU CGAUAUUGAUUUAAAAGAAAUCAGCUCUAAAACACUGGGACUUGAU AAGCUUAAUGUCCAAGAUGCCUACACCCCGAAAGAAACUGCUGUAAC CGUUGAUAAAACUACCUAUAAAAAUGGUACAGAUCCUAUUACAGCCC AGAGCAAUACUGAUAUCCAAACUGCAAUUGGCGGUGGUGCAACGGGG GUUACUGGGGCUGAUAUCAAAUUUAAAGAUGGUCAAUACUAUUUAG AUGUUAAAGGCGGUGCUUCUGCUGGUGUUUAUAAAGCCACUUAUGA UGAAACUACAAAGAAAGUUAAUAUUGAUACGACUGAUAAAACUCCG UUGGCAACUGCGGAAGCUACAGCUAUUCGGGGAACGGCCACUAUAAC CCACAACCAAAUUGCUGAAGUAACAAAAGAGGGUGUUGAUACGACCA CAGUUGCGGCUCAACUUGCUGCAGCAGGGGUUACUGGCGCCGAUAAG GACAAUACUAGCCUUGUAAAACUAUCGUUUGAGGAUAAAAACGGUA AGGUUAUUGAUGGUGGCUAUGCAGUGAAAAUGGGCGACGAUUUCUA UGCCGCUACAUAUGAUGAGAAAACAGGUGCAAUUACUGCUAAAACCA CUACUUAUACAGAUGGUACUGGCGUUGCUCAAACUGGAGCUGUGAAA UUUGGUGGCGCAAAUGGUAAAUCUGAAGUUGUUACUGCUACCGAUG GUAAGACUUACUUAGCAAGCGACCUUGACAAACAUAACUUCAGAACA GGCGGUGAGCUUAAAGAGGUUAAUACAGAUAAGACUGAAAACCCAC UGCAGAAAAUUGAUGCUGCCUUGGCACAGGUUGAUACACUUCGUUCU GACCUGGGUGCGGUUCAGAACCGUUUCAACUCCGCUAUCACCAACCU GGGCAAUACCGUAAAUAACCUGUCUUCUGCCCGUAGCCGUAUCGAAG AUUCCGACUACGCAACCGAAGUCUCCAACAUGUCUCGCGCGCAGAUU CUGCAGCAGGCCGGUACCUCCGUUCUGGCGCAGGCGAACCAGGUUCC GCAAAACGUCCUCUCUUUACUGCGUUGAUAAUAGGCUGGAGCCUCGG UGGCCAUGCUUCUUGCCCCUUGGGCCUCCCCCCAGCCCCUCCUCCCCU UCCUGCACCCGUACCCCCGUGGUCUUUGAAUAAAGUCUGAGUGGGCG GCAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAUCUAG

TABLE 66  Flagellin Amino Acid Sequences SEQ ID Name Sequence NO: ORF MAQVINTNSLSLLTQNNLNKSQSALGTAIERLSSGLRINSAKDDAAGQAIA 420 Sequence, NRFTANIKGLTQASRNANDGISIAQTTEGALNEINNNLQRVRELAVQSANG AA TNSQSDLDSIQAEITQRLNEIDRVSGQTQFNGVKVLAQDNTLTIQVGANDG ETIDIDLKEISSKTLGLDKLNVQDAYTPKETAVTVDKTTYKNGTDPITAQSN TDIQTAIGGGATGVTGADIKFKDGQYYLDVKGGASAGVYKATYDETTKK VNIDTTDKTPLATAEATAIRGTATITHNQIAEVTKEGVDTTTVAAQLAAAG VTGADKDNTSLVKLSFEDKNGKVIDGGYAVKMGDDFYAATYDEKTGAIT AKTTTYTDGTGVAQTGAVKFGGANGKSEVVTATDGKTYLASDLDKHNFR TGGELKEVNTDKTENPLQKIDAALAQVDTLRSDLGAVQNRFNSAITNLGN TVNNLSSARSRIEDSDYATEVSNMSRAQILQQAGTSINVLAQANQVPQNVL SLLR Flagellin- MAQVINTNSLSLLTQNNLNKSQSALGTAIERLSSGLRINSAKDDAAGQAIA 421 GS linker- NRFTANIKGLTQASRNANDGISIAQTTEGALNEINNNLQRVRELAVQSANS circumspor TNSQSDLDSIQAEITQRLNEIDRVSGQTQFNGVKVLAQDNTLTIQVGANDG ozoite ETIDIDLKQINSQTLGLDTLNVQQKYKVSDTAATVTGYADTTIALDNSTFK protein ASATGLGGTDQKIDGDLKFDDTTGKYYAKVTVTGGTGKDGYYEVSINVD (CSP) KTNGEVTLAGGATSPLTGGLPATATEDVKNVQVANADLTEAKAALTAAG VTGTASINVVKMSYTDNNGKTIDGGLAVKVGDDYYSATQNKDGSISINTT KYTADDGTSKTALNKLGGADGKTEVVSIGGKTYAASKAEGHNFKAQPDL AEAAATTTENPLQKIDAALAQVDTLRSDLGAVQNRFNSAITNLGNTVNNL TSARSRIEDSDYATEVSNMSRAQILQQAGTSINVLAQANQVPQNVLSLLRG GGGSGGGGSMMAPDPNANPNANPNANPNANPNANPNANPNANPNANPN ANPNANPNANPNANPNANPNANPNANPNANPNANPNANPNANPNKNNQ GNGQGHNMPNDPNRNVDENANANNAVKNNNNEEPSDKHIEQYLKKIKNS ISTEWSPCSINVTCGNGIQVRIKPGSANKPKDELDYENDIEKKICKMEKCSSI NVFNVVNS Flagellin- MMAPDPNANPNANPNANPNANPNANPNANPNANPNANPNANPNANPNA 422 RPVT NPNANPNANPNANPNANPNANPNANPNANPNANPNKNNQGNGQGHNMP linker- NDPNRNVDENANANNAVKNNNNEEPSDKHIEQYLKKIKNSISTEWSPCSIN circumspor VTCGNGIQVRIKPGSANKPKDELDYENDIEKKICKMEKCSSINVFNVVNSR ozoite PVT MAQVINTNSLSLLTQNNLNKSQSALGTAIERLSSGLRINSAKDDAAGQ protein AIANRFTANIKGLTQASRNANDGISIAQTTEGALNEINNNLQRVRELAVQS (CSP) ANSTNSQSDLDSIQAEITQRLNEIDRVSGQTQFNGVKVLAQDNTLTIQVGA NDGETIDIDLKQINSQTLGLDTLNVQQKYKVSDTAATVTGYADTTIALDNS TFKASATGLGGTDQKIDGDLKFDDTTGKYYAKVTVTGGTGKDGYYEVSIN VDKTNGEVTLAGGATSPLTGGLPATATEDVKNVQVANADLTEAKAALTA AGVTGTASINVVKMSYTDNNGKTIDGGLAVKVGDDYYSATQNKDGSISIN TTKYTADDGTSKTALNKLGGADGKTEVVSIGGKTYAASKAEGHNFKAQP DLAEAAATTTENPLQKIDAALAQVDTLRSDLGAVQNRFNSAITNLGNTVN NLTSARSRIEDSDYATEVSNMSRAQILQQAGTSINVLAQANQVPQNVLSLL R

TABLE 67 Signal Peptides SEQ ID Description Sequence NO: HuIgG_(k) signal peptide METPAQLLFLLLLWLPDTTG 423 IgE heavy chain epsilon MDWTWILFLVAAATRVHS 424 −1 signal peptide Japanese encephalitis  MLGSNSGQRVVFTILLLLVA 425 PRM signal sequence PAYS VSINVg protein  MKCLLYLAFLFIGVNCA 426 signal sequence Japanese encephalitis  MWLVSLAIVTACAGA 427 JEV signal sequence

EQUIVALENTS

Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the disclosure described herein. Such equivalents are intended to be encompassed by the following claims.

All references, including patent documents, disclosed herein are incorporated by reference in their entirety. 

1-147. (canceled)
 148. A method of inducing an immune response in a subject, the method comprising administering to the subject in an amount effective a Chikungunya virus (CHIKV) vaccine comprising a ribonucleic acid (RNA) polynucleotide having an open reading frame encoding a CHIKV polyprotein comprising C protein, E1 protein, E2 protein and E3 protein formulated in a lipid nanoparticle that comprises a molar ratio of 20-60% ionizable cationic lipid, 5-25% non-cationic lipid, 25-55% sterol, and 0.5-15% PEG-modified lipid, in an effective amount to produce an antigen-specific immune response in the subject.
 149. The method of claim 148, wherein the antigen specific immune response comprises a T cell response or a B cell response.
 150. The method of claim 148, wherein the subject is administered a single dose of the vaccine.
 151. The method of claim 148, wherein the subject is administered a booster dose of the vaccine.
 152. The method of claim 148, wherein the vaccine is administered to the subject by intradermal injection or intramuscular injection.
 153. The method of claim 148, wherein an anti-CHIKV polyprotein antibody titer produced in the subject is increased by at least 1 log and/or at least 2 times relative to a control. 154.-156. (canceled)
 157. The method of claim 153, wherein the control is selected from an anti-CHIKV polyprotein antibody titer produced in a subject who has not been administered a vaccine against the virus, an anti-CHIKV polyprotein antibody titer produced in a subject who has been administered a live attenuated vaccine or an inactivated vaccine against the virus, an anti-CHIKV polyprotein antibody titer produced in a subject who has been administered a recombinant protein vaccine or purified protein vaccine against the virus, and an anti-CHIKV polyprotein antibody titer produced in a subject who has been administered a VLP vaccine against the virus. 158.-185. (canceled)
 186. The method of claim 148, wherein the CHIKV polyprotein further comprises 6K protein.
 187. The method of claim 148, wherein the RNA polynucleotide further encodes 5′ terminal cap, 7mG(5′)ppp(5′)NlmpNp.
 188. The method of claim 148, wherein at least 80% of the uracil in the open reading frame have a chemical modification selected from N1-methyl-pseudouridine or N1-ethyl-pseudouridine.
 189. The method of claim 188, wherein the chemical modification is in the 5-position of the uracil.
 190. The method of claim 148, wherein the efficacy of the vaccine is at least 70%, relative to unvaccinated subjects, following a single dose of the vaccine.
 191. The method of claim 148, wherein the effective amount is sufficient to produce detectable levels of CHIKV polyprotein as measured in serum of the subject at 1-72 hours post administration.
 192. The method of claim 148, wherein the effective amount is sufficient to produce a 1,000-10,000 neutralization titer produced by neutralizing antibody against the CHIKV polyprotein as measured in serum of the subject at 1-72 hours post administration.
 193. The method of claim 148, wherein the effective amount is a total dose of 25 μg-400 μg.
 194. The method of claim 148, wherein the ionizable cationic lipid comprises the following compound:


195. The method of claim 149, wherein the CHIKV polyprotein comprises an amino acid sequence that has at least 90% identity to the amino acid sequence identified by SEQ ID NO:
 408. 196. The method of claim 195, wherein the CHIKV polyprotein comprises the amino acid sequence identified by SEQ ID NO:
 408. 197. The method of claim 149, wherein the RNA polynucleotide has an open reading frame that comprises a nucleotide sequence that has at least 90% identity to the RNA sequence identified by SEQ ID NO:
 401. 198. The method of claim 197, wherein the RNA polynucleotide has an open reading frame that comprises the nucleotide sequence identified by SEQ ID NO:
 401. 199. The method of claim 148, wherein the CHIKV vaccine further comprises a RNA polynucleotide having an open reading frame encoding a Zika virus (ZIKV) polypeptide.
 200. The method of claim 199, wherein the ZIKV polypeptide comprises a ZIKV prME protein.
 201. The method of claim 200, wherein the ZIKV prME protein comprises an amino acid sequence that has at least 90% identity to the amino acid sequence identified by SEQ ID NO:
 203. 202. The method of claim 200, wherein the RNA polynucleotide having an open reading frame encoding a ZIKV prME protein comprises a nucleotide sequence that has at least 90% identity to the nucleotide sequence identified by SEQ ID NO:
 139. 203. The method of claim 148, wherein the CHIKV vaccine further comprises a RNA polynucleotide having an open reading frame encoding a Dengue virus (DENV) polypeptide.
 204. The method of claim 203, wherein the DENV polypeptide comprises aDENV prME protein.
 205. The method of claim 204, wherein the DENV prME protein comprises an amino acid sequence that has at least 90% identity to the amino acid sequence identified by SEQ ID NO:
 267. 206. The method of claim 204, wherein the RNA polynucleotide having an open reading frame encoding DENV prME protein comprises a nucleotide sequence that has at least 90% identity to the nucleotide sequence identified by SEQ ID NO:
 248. 